Search

A novel sample extraction method using an ISOLUTE PLD+ protein and phospholipid removal column was developed for simultaneous quantification of 20 psychoactive drugs, including antidepressants, antipsychotics, sedative-hypnotics, and amphetamines, in postmortem whole blood samples by liquid chromatography-tandem mass spectrometry. The method showed improvement in extract cleanliness compared with traditional protein precipitation and the QuEChERS extraction method. The method was validated for all analytes; the calibration curves showed good linearity, with r

Objective Melanocytes are widely distributed in the skin and often in oral mucosa but normally do not exist in bone tissue. The aim of this study was to compare the existence of melanocytes in odontogenic keratocysts (OKCs) and radicular cysts (RCs) and to clarify the difference in origin between those cysts. Methods One hundred and ten cases of OKCs and RCs were used. In OKC, 88 cases showed sporadic type (SPO), and 22 cases involved basal cell nevus syndrome (BCNS). OKC samples were divided into 54 cases in the juvenile group (0-29 years old) and 56 cases in the advanced group (30-70 years old). We researched these cysts with Melan-A and HMB45 immunohistochemical and Schmorl's reaction staining. Melanocytes were detected using Melan-A and HMB45 immunohistochemical stainings, and melanin pigmentation was detected using Schmorl's method. Results Melanocytes and melanin pigmentation were shown in OKC, and neither melanin pigmentation nor melanocytes existed in RC. The positive rate of Schmorl's reaction, Melan-A, and HMB45 staining were significantly higher in the juvenile group than in the advanced group. These rates were also higher in BCNS than in SPO. Conclusions Our data raise the important possibility that the origin of OKC epithelium differs from RC by the expression of melanocytes and melanin pigmentation. Our findings also underscore the fact that the origin of OKC differs between younger and older patients. It is suggested that melanin pigmentation and melanocytes expression may assist in the classification of odontogenic cysts in the future.

Oral potentially malignant disorders are associated with the development of oral squamous cell carcinoma (OSCC). Most OSCCs are diagnosed via histopathology as oral epithelial dysplasia (OED), but the histologic diagnostic criteria remain non-uniform. Accordingly, the establishment of a diagnostic marker to assist in diagnosis could contribute towards cancer prevention. Melanoma inhibitory activity (MIA) and MIA2 are involved in tumor growth, invasion, and lymph node metastasis in various malignancies. The purpose of this study was to clarify the usefulness of MIA and MIA2 as diagnostic markers of oral mucosal lesions. The expression of MIA and MIA2 was analyzed immunohistochemically in 100 specimens (10 specimens with normal oral mucosa (NOM) and 30 specimens each with low-grade epithelial dysplasia (LED), high-grade epithelial dysplasia (HED), and OSCC). Immunohistochemical results were evaluated based on the Allred scoring system. Cytoplasmic expression of MIA and MIA2 increased in the order of LED, HED, and OSCC. All NOM specimens were negative for cytoplasmic expression. Significant differences were observed between the groups (NOM vs. HED, p &lt, 0.05, NOM vs. OSCC, p &lt, 0.001). These results demonstrate that MIA and MIA2 are expressed in the oral mucosa within early neoplastic lesions and suggest that MIA and MIA2 are useful novel immunohistochemical markers for discriminating between normal tissue and OED.

Titanium implants are the standard therapeutic option when restoring missing teeth and reconstructing fractured and/or diseased bone. However, in the 30 years since the advent of micro-rough surfaces, titanium&rsquo, s ability to integrate with bone has not improved significantly. We developed a method to create a unique titanium surface with distinct roughness features at meso-, micro-, and nano-scales. We sought to determine the biological ability of the surface and optimize it for better osseointegration. Commercially pure titanium was acid-etched with sulfuric acid at different temperatures (120, 130, 140, and 150 &deg, C). Although only the typical micro-scale compartmental structure was formed during acid-etching at 120 and 130 &deg, C, meso-scale spikes (20&ndash, 50 &mu, m wide) and nano-scale polymorphic structures as well as micro-scale compartmental structures formed exclusively at 140 and 150 &deg, C. The average surface roughness (Ra) of the three-scale rough surface was 6&ndash, 12 times greater than that with micro-roughness only, and did not compromise the initial attachment and spreading of osteoblasts despite its considerably increased surface roughness. The new surface promoted osteoblast differentiation and in vivo osseointegration significantly, regression analysis between osteoconductivity and surface variables revealed these effects were highly correlated with the size and density of meso-scale spikes. The overall strength of osseointegration was the greatest when the acid-etching was performed at 140 &deg, C. Thus, we demonstrated that our meso-, micro-, and nano-scale rough titanium surface generates substantially increased osteoconductive and osseointegrative ability over the well-established micro-rough titanium surface. This novel surface is expected to be utilized in dental and various types of orthopedic surgical implants, as well as titanium-based bone engineering scaffolds.

Objective The present study was carried out to histologically, immunohistochemically, and histometrically investigate the effect of low-level laser irradiation by a diode laser on the growth of the rat mandibular condyle in organ culture. Methods The mandibular condyles were removed from rats’ embryos on the 21st day and then cultured for 8 days. The 20 cultured condyles were divided into 4 groups: F−L− group (serumless BGJb); F+L− group (serumless BGJb with bFGF 100 ng/mL); F−L+ group (serumless BGJb and laser irradiation); and F+L+ group (serumless BGJb with bFGF 100 ng/mL and laser irradiation). The laser irradiation (output 1.3 W/cm2 and wavelength 633 nm) was performed on the cultured condyle for 30 seconds every 24 hours for 5 days. Immunohistochemically, PCNA was used to analyze cell proliferation. Results The mesenchymal cells in the layer of differentiation and hypertrophy in the F+L+ group showed a marked increase in cell number and the size of the condyle. The positive cell number of PCNA in F+L− and F+L+ group was significantly higher compared with the control group. Conclusions Our data showed that low-level laser irradiation promotes cell proliferation in the cultured condyle same as in bFGF addition. It is suggested that low-level laser irradiation by a diode laser (wavelength 633 nm) stimulated the proliferation of mesenchymal cells, which resulted in an increase in the number of chondrocytes in the layers of differentiation and hypertrophy by leading to the enlargement of the condylar size.

Objectives Melanocytes are pigmented-producing cells and derived from the neural crest. Melanin pigmentation is widely distributed in the skin and often in oral mucosa, but normally not existing in bone tissue. However, melanin pigmentation is detected on rare occasions with odontogenic lesions in the jaw bones, especially odontogenic keratocysts (OKC). Moreover, development of the tooth germ is originated from the neural crest, but elimination or expression of neural crest cells with odontogenic lesions is not obvious. The present study aimed to consider the origins of OKC based on the distribution of melanocytes in OKC. Findings One hundred and ten OKC were used. Eighty-eight cases showed sporadic type (SPO), and 22 cases involved basal cell nevus syndrome (BCNS). All samples were divided into 54 cases of juvenile group (0-29 years old) and 56 cases of advanced group (30-70 years old). Melanocytes were detected using Melan-A and HMB45 immunohistochemical stainings, and melanin pigmentation was detected using Schmorl's method. The positive rate of Schmorl's reaction, Melan-A and HMB45 staining were significantly higher in juvenile group than advanced group. These rates were also higher in BCNS than SPO. Conclusions Compare to juvenile and advanced groups, Melan-A and HMB45 positive rates were high in juvenile group. It is evident from these findings that the origin of OKC in juvenile group was different from advanced one. It means that the cyst epithelium in juvenile group originated from neural crest cell with melanocytes, and advanced one arose from odontogenic epithelium without melanocyte, for examples epithelial rest of Malassez.

The expression of Notch in 30 cases of pleomorphic adenoma was examined by immunohistochemistry. Comparing the results of our study with previous literatures, from the partial CK7 expression and substantial Notch expression in ductal epithelial cells as well as the Notch expression in solid tumor nests, it can be inferred that Notch is involved in cell differentiation. CK13 expression was observed in cells undergoing squamous metaplasia and Notch expression was seen in the nucleus of basal and squamous cells. The intense Notch expression in basal cells and weak expression in squamous cells suggests that Notch is involved in the differentiation from basal to squamous cell. Moreover, the loss of nuclear expression on the inner layer would signify that differentiation is about to end or has been terminated. Notch was expressed in the cytoplasm of cartilage cells and in the cell membrane of mucous cells but not in the nucleus indicating that differentiation has been concluded. Notch involvement is suspected in cell differentiation in areas showing ductal structures and squamous metaplasia. In summary, Notch is involved in cell differentiation of ductal cells in PA. Nuclear expression was shown in tumor cells in solid nests and surrounding structures. Moreover, Notch is expressed by basal cells undergoing squamous metaplasia suggesting the participation of Notch in cell differentiation in PA.

There are well known that Wnt signaling was some roles of cell differentiation at the development tissues, especially the oral and maxillofacial regions of some developmental stages. Therefore, to determine Wnt signaling in the pleomorphic adenoma tissues, we examined. The expression of Wnt1 and β-catenin as well as the distribution of various cytoskeletal proteins CK7 and CK13 was examined in 30 cases of pleomorphic adenoma by immunohistochemistry. Wnt1 was detected in almost all tumor cells. The peripheral columnar cells in squamous metaplasia and small cuboidal cells in duct-like structures were strongly positive to Wnt1. Although β-catenin was clearly localized on the cell membrane of tumor cells, nuclear translocation was observed in small cuboidal cells and in some basaloid cells. The immunofluorescent staining pattern of Wnt1 and CK7 as well as Wnt1 and CK13 was consistent with IHC results. Thus, in pleomorphic adenoma, Wnt is involved in tumor cell differentiation of peripheral columnar cells forming solid nests and small peripheral columnar cells forming duct-like structures. Moreover, among the three currently known Wnt pathways, β-catenin is the suggested pathway working during cell differentiation. Furthermore, peripheral columnar cells in solid tumor nests and in squamous metaplasia are governed by another Wnt pathway other than β-catenin. Therefore, Wnt signaling through β-catenin pathway may be involved in the ‘mixed’ differentiation characteristic of pleomorphic adenoma although another pathway may also be possibly working in other parts of the tumor tissue., 2014

Bone regeneration often requires cues from osteogenesis-inducing factors for successful outcome. N-acetyl cysteine (NAC), an anti-oxidant small molecule, possibly modulates osteoblastic differentiation. This study investigated the potential of NAC as an osteogenesis-enhancing molecule in vitro and in vivo. Various concentrations of NAC (0, 2.5, 5.0, and 10 mM) were added to rat bone marrow stromal cell or osteoblastic cell culture in media with or without dexamethasone. The results showed marked enhancement of alkaline phosphatase activity and mineralized matrix formation together with consistent upregulation of bone-related gene markers such as collagen I, osteopontin, and osteocalcin in the osteoblastic culture with addition of 2.5 or 5.0 mM NAC regardless of the presence of dexamethasone. Micro-CT-based analysis and histological observation revealed that addition of NAC to a collagenous sponge implanted in a critical size cortical bone defect (3.0 mm × 5.0 mm) in rat femur yielded acceleration and completion of defect closure, with thick, compact, and contiguous bone after 6 weeks of healing. In contrast, with sponge alone, only sparse and incomplete bone regeneration was observed during the matching healing period. These results indicate that NAC can function as an osteogenesis-enhancing molecule to accelerate bone regeneration by activating differentiation of osteogenic lineages.

The expression of HSP27 and some CKs were examined the 40 cases of typical solid/multicystic ameloblastoma using immunohistochemical techniques. In order to examine the relevance of HSP in cell differentiation, we focused on the cytoskeletal expression of CK. CK19 is a marker of typical odontogenic epithelium widely observed in follicular and plexiform types of ameloblastomas. Since staining with CK14 is one of the measures of the differentiation potential of squamous cells and is extensively expressed in both follicular and plexiform types, it implies that squamous differentiation of each type can occur. CK8 was strongly detected in tumor nests in plexiform type but weakly detected in follicular type. It was considered that the expression of HSP27 in plexiform type correlated with the expression of CK8 suggesting that HSP27 might have regulated the expression of CK8.

近年、Electrochemotherapy (ECT)はパルス状電圧によって抗腫瘍剤を腫瘍局所の細胞内に取り込ませる腫瘍の治療方法の一つとして注目されている。しかし、その治療方法はまだ確立されていない。そこで本研究では、口腔領域に多く発生する扁平上皮癌の実験動物モデルを使用し、パルス状低電圧負荷によるブレオマイシン(BLM)の抗腫瘍効果について検索を行った。実験ではヒト子宮頚部癌扁平上皮癌細胞(Caski)を培養後にヌードマウスに移植し、その後、腫瘍移植部に対してECTの処置を行った。その結果、D+E+群（薬剤投与電気穿孔群）では腫瘍体積の減少が認められた。また、数例では腫瘍の消失が認められた。これらの結果から、BLM投与とともにECTの処置を加えることで、薬剤の抗腫瘍作用は増強されることが判明した。本研究の結果から、ECTは腫瘍の治療において、安全で効果的な新しい治療法として有用であることが示唆された。

The objectives of this in vitro study were to determine whether the commercial collagen material used in bone augmentation procedures induces oxidative stress-mediated adverse effects on the viability and function of osteoblasts and to determine whether N-acetyl cysteine (NAC), an antioxidant amino acid derivative, can alleviate these effects.Commercial collagen sponge (Collaplug) and membrane (BioGide) were treated with NAC. Rat calvaria-derived osteoblasts were directly seeded on these materials with or without NAC pretreatment. Cytotoxic evaluation was performed by flowcytometric cell viability assay, confocal laser microscopic analysis of attached cell morphology and reactive oxygen species (ROS) localization, and alkaline phosphatase staining.Cell viability was less than 40% on both collagen sponge and membrane 24 hours after seeding and increased to 50% with NAC pretreatment. Cell death was characterized by apoptosis. Colonization of attached cells was sparse on the untreated sponge and membrane on day 1, and the cells were round, small, and filled with intense and closely packed intracellular ROS. In contrast, NAC-pretreated material had dense cell colonies consisting of well-spread osteoblasts and fully developing cytoskeleton and cellular processes with little ROS generation. On day 7 of culture, NAC-pretreated collagen sponge and membrane yielded an expanded alkaline phosphatase-positive area occupying 60% and 80% of the surface area, respectively, whereas the untreated collagen materials had limited alkaline phosphatase activity (7% or less).Commercial collagen sponge and membrane induced considerable cell death, impaired initial function, and generated extraordinary intracellular ROS in attached osteoblasts, whereas NAC pretreatment substantially ameliorated these effects. The potential benefits of NAC's detoxifying capacity on bone regeneration using collagen matrix materials in an animal model should be confirmed with further study.

This study examines the cytotoxicity of bone cement extract to osteoblasts and the potential detoxification and restoration of osteoblastic function by an antioxidant amino acid, N-acetyl cysteine (NAC). The osteoblastic cells derived from rat femurs were cultured with extract from polymethyl methacrylate (PMMA)-based bone cement. The calcein and ethidium homodimer staining of the cells after 24-h incubation showed that 23.0% of the cells were dead in the culture with bone cement extract, while the addition of 5 mM NAC into the culture reduced the percentage to 4.3%. Annexin V and propidium iodide-based flow cytometric analysis also revealed that the apoptotic cells present at 15.8% in the culture with bone cement extract was reduced to 2.4% in the culture cotreated with bone cement extract and NAC. Severely suppressed alkaline phosphatase activity and matrix mineralization in the culture with bone cement extract (reduced to 10% and 5%, respectively, compared with the control culture) were restored to a normal level when treated with 5 mM NAC. The bone cement extract-induced, downregulated expression of osteoblastic genes, such as alkaline phosphatase, collagen I, and osteocalcin, was also restored to the baseline level by cotreatment with NAC. The data indicated that the addition of NAC into acrylic bone cement extract remarkably ameliorated the cytotoxicity to osteoblasts and restored their phenotype and function to a biologically significant degree, suggesting the potential usefulness of NAC in developing more biocompatible acrylic bone cement.

Biological tissues involve hierarchical organizations of structures and components. We created a micropit-and-nanonodule hybrid topography of TiO(2) by applying a recently reported nanonodular self-assembly technique on acid-etch-created micropit titanium surfaces. The size of the nanonodules was controllable by changing the assembly time. The created micro-nano-hybrid surface rendered a greater surface area and roughness, and extensive geographical undercut on the existing micropit surface and resembled the surface morphology of biomineralized matrices. Rat bone marrow-derived osteoblasts were cultured on titanium disks with either micropits alone, micropits with 100-nm nodules, micropits with 300-nm nodules, or micropits with 500-nm nodules. The addition of nanonodules to micropits selectively promoted osteoblast but not fibroblast function. Unlike the reported advantages of microfeatures that promote osteoblast differentiation but inhibit its proliferation, micro-nano-hybrid topography substantially enhanced both. We also demonstrated that these biological effects were most pronounced when the nanonodules were tailored to a diameter of 300nm within the micropits. An implant biomechanical test in a rat femur model revealed that the strength of bone-titanium integration was more than three times greater for the implants with micropits and 300-nm nanonodules than the implants with micropits alone. These results suggest the establishment of functionalized nano-in-microtitanium surfaces for improved osteoconductivity, and may provide a biomimetic micro-to-nanoscale hierarchical model to study the nanofeatures of biomaterials.

Introduction Dental pulp stem cells (DPSCs) are mesenchymal stem cells located in dental pulp and are thought to be a potential source for cell therapy since DPSCs can be easily obtained from teeth extracted for orthodontic reasons. Obtained DPSCs can be cryopreserved until necessary and thawed and expanded when needed. The aim of this study is to evaluate the therapeutic potential of DPSC transplantation for diabetic polyneuropathy. Methods DPSCs isolated from the dental pulp of extracted incisors of Sprague–Dawley rats were partly frozen in a −80 °C freezer for 6 months. Cultured DPSCs were transplanted into the unilateral hindlimb skeletal muscles 8 weeks after streptozotocine injection and the effects of DPSC transplantation were evaluated 4 weeks after the transplantation. Results Transplantation of DPSCs significantly improved the impaired sciatic nerve blood flow, sciatic motor/sensory nerve conduction velocity, capillary number to muscle fiber ratio and intra-epidermal nerve fiber density in the transplanted side of diabetic rats. Cryopreservation of DPSCs did not impair their proliferative or differential ability. The transplantation of cryopreserved DPSCs ameliorated sciatic nerve blood flow and sciatic nerve conduction velocity as well as freshly isolated DPSCs. Conclusions We demonstrated the effectiveness of DPSC transplantation for diabetic polyneuropathy even when using cryopreserved DPSCs, suggesting that the transplantation of DPSCs could be a promising tool for the treatment of diabetic neuropathy.

Atelocollagen sponges incorporating polyhedra encapsulating bone morphogenetic protein 2 (BMP-2) were implanted into lateral bone defects in the mandible. Half of the bone defects on the left side were treated with atelocollagen sponges containing 1.8 × 107 BMP-2 polyhedra, and half were treated with sponges containing 3.6 × 106 BMP-2 polyhedra. As controls, we treated the right-side bone defects in each animal with an atelocollagen sponge containing 5 µg of recombinant human BMP-2 (rhBMP-2) or 1.8 × 107 empty polyhedral. After a healing period of six months, whole mandibles were removed for micro-computed tomography (CT) and histological analyses. Micro-CT images showed that more bone had formed at all experimental sites than at control sites. However, the density of the new bone was not significantly higher at sites with an atelocollagen sponge containing BMP-2 polyhedra than at sites with an atelocollagen sponge containing rhBMP-2 or empty polyhedra. Histological examination confirmed that the BMP-2 polyhedra almost entirely replaced the atelocollagen sponges and connected the original bone with the regenerated bone. These results show that the BMP-2 delivery system facilitates the regeneration of new bone in the mandibular alveolar bone ridge and has an advance in the technology of bone regeneration for implant site development.

In the present study, the effects of both growth factor TGF- α and carcinogen Trp-P-2 on the development of organ-cultured ddY mouse tooth-germ which was transplanted into the subcutaneous tissue of nude mice were investigated histologically, immnohistochemically (PCNA) and histometrically in 5 groups of mice, namely Group A (Control), Group B (DMSO), Group C (TGF-α), Group D (Trp-P-2) and Group E (TGF-α + Trp-P-2). Histologically, in Groups C, D and E, marked proliferation of odontogenic epithelial cells was observed. In Groups D and E, keratinization of the odontogenic epithelial cells was found, but it was not observed in Group C. Moreover, in Group E, a cyst-like space was observed in the area of odontogenic epithelial cells. Immunohistochemically, in Groups C, D and E, PCNA-positivities of the odontogenic epithelial cells were significantly higher than those of Group A. Histometrically, in Groups C and E, the size of the tooth germ was significantly larger than that of Group A. The results of the present study suggest that proliferation, keratinization, cyst-like space formation of odontogenic epitherial cells of transplanted organ-cultured tooth germs in mice were promoted by growth factor TGF- α and carcinogen Trp-P-2.

Purpose: Peri-implant osteogenesis is reported to be impaired in patients with diabetes. The current study tested the hypothesis that ultraviolet (UV) treatment of titanium, or photofunctionalization, is able to mitigate the impaired osseointegration associated with type 2 diabetes. Materials and Methods: Untreated and photofunctionalized titanium implants were placed into the femurs of genetically modified rats with a close phenotypic resemblance to human type 2 diabetes, as characterized by late-onset hyperglycemia and obesity. Implants were photofunctionalized with UV light for 15 minutes immediately before placement. The strength of osseointegration was evaluated using a biomechanical push-in test, and the tissue-implant interface was examined using scanning electron microscopy and energy-dispersive spectroscopy. Results: Photofunctionalization converted implants from hydrophobic to superhydrophilic. Photofunctionalizationinduced hemophilicity was also confirmed during surgery. The strength of osseointegration of photofunctionalized implants was significantly greater than that of untreated implants, by 1.8 and 3 times, at weeks 2 and 4 of healing, respectively. Osseointegration of photofunctionalized implants in diabetic animals was even stronger than that of untreated implants placed in normal animals throughout the healing period. Photofunctionalized implants placed in diabetic rats were extensively covered with calcium- and phosphorusrich tissue that masked the titanium signal. Conclusion: Photofunctionalization accelerated and enhanced levels of osseointegration and overcame impaired osseointegration in a rat model of type 2 diabetes. Further prospective studies are warranted to establish the clinical efficacy of photofunctionalization in patients with diabetes.

The aim of this study was to show the effectiveness of combining calcium phosphate cement and gelatin powders to promote bone regeneration in the canine mandible. We mixed gelatin powders with calcium phosphate cement to create a macroporous composite. In four beagle dogs, two saddle-type bone defects were created on each side of the mandible, and calcium phosphate cement alone or calcium phosphate cement containing composite gelatin powders was implanted in each of the defects. After a healing period of six months, mandibles were removed for µCT and histological analyses. The µCT and histological analyses showed that at experimental sites at which calcium phosphate cement alone had been placed new bone had formed only around the periphery of the residual calcium phosphate cement and that there had been little or no ingrowth into the calcium phosphate cement. On the other hand, at experimental sites at which calcium phosphate cement containing composite gelatin powders had been placed, we observed regenerated new bone in the interior of the residual calcium phosphate cement as well as around its periphery. The amount of resorption of calcium phosphate cement and bone regeneration depended on the mixing ratio of gelatin powders to calcium phosphate cement. New bone replacement was significantly better in the sites treated with calcium phosphate cement containing composite gelatin powders than in those treated with calcium phosphate cement alone.

Bombyx mori cypovirus is a major pathogen which causes significant losses in silkworm cocoon harvests because the virus particles are embedded in micrometer-sized protein crystals called polyhedra and can remain infectious in harsh environmental conditions for years. But the remarkable stability of polyhedra can be applied on slow-release carriers of cytokines for tissue engineering. Here we show the complete healing in critical-sized bone defects by bone morphogenetic protein-2 (BMP-2) encapsulated polyhedra. Although absorbable collagen sponge (ACS) safely and effectively delivers recombinant human BMP-2 (rhBMP-2) into healing tissue, the current therapeutic regimens release rhBMP-2 at an initially high rate after which the rate declines rapidly. ACS impregnated with BMP-2 polyhedra had enough osteogenic activity to promote complete healing in critical-sized bone defects, but ACS with a high dose of rhBMP-2 showed incomplete bone healing, indicating that polyhedral microcrystals containing BMP-2 promise to advance the state of the art of bone healing.

The aim of this study was to qualitatively evaluate a poly(lactic acid-co-glycolic acid-co-ε-caprolactone) (PLGC) membrane as a barrier for guided bone regeneration in the canine mandible and to compare it to a nonresorbable polytetrafluoroethylene (PTFE) membrane.Two wedge-shaped bone defects were created bilaterally in the mandibles of 12 beagle dogs. The bone defects in the left mandible were divided into three groups and treated as follows: PLGC membrane alone, PLGC membrane plus autogenous cortical bone chips, and titanium-reinforced expanded PTFE (TR-PTFE) membrane. The bone defects in the right mandible of each animal were left without membranes as a control. Computed tomography (CT) was performed at 3 and 6 months postoperative to evaluate bone regeneration. After a healing period of 6 months, the mandibles were removed en bloc for micro-CT and histologic analyses.CT analyses at 3 and 6 months showed that there was significantly more bone augmentation at all experimental sites than at the control sites. The volume of bone at defect sites covered with TR-PTFE was significantly greater than at defect sites covered with PLGC membrane with or without autogenous cortical bone. Micro-CT measurements showed that the volume of new bone formed at sites covered with TR-PTFE was significantly greater than at sites covered with PLGC membrane. However, the density of new bone was significantly higher at sites covered with PLGC membrane, with or without cortical bone, than at sites covered with TR-PTFE. Histologic analysis verified the presence of well-vascularized loose connective tissue in the pores of the PLGC membrane.Compared to TR-PTFE, the macroporous bioresorbable PLGC membrane did not significantly increase the amount of new bone in defect sites, but it facilitated the regeneration of mature bone.