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Introduction: The lips fulfill various critical physiological roles besides being viewed as a fundamental aesthetic feature contributing to the perception of health and beauty. Therefore, any lip injury, abnormality, or congenital malformation, such as cleft lip, needs special attention in order to restore proper lip function and aesthetics. To achieve this goal, a better understanding of the complex lip anatomy, function, and biology is required, which can only be provided by basic research endeavors. However, the current lack of clinically relevant human lip cells and three-dimensional in vitro lip models, capable of replacing ethically questionable animal experimentations, represents a significant limitation in this area of research. Methods: To address these limitations, we aimed to pioneer the introduction of immortalized healthy lip- and cleft lip-derived keratinocytes. Primary keratinocytes were isolated from patients' samples and immortalized by introducing the catalytic domain of telomerase, combined with the targeted knockdown of the cell cycle inhibitor gene, p16 INK4A . We then focused on validating the newly established cell lines by comparing their genetic stability and key phenotypic features with their primary keratinocyte counterparts. Results: The newly established immortalized keratinocyte cell lines demonstrated genetic stability and preserved the main phenotypic characteristics of primary keratinocytes, such as cellular morphology and differentiation capacity. Three-dimensional lip models, generated using these cell lines, proved to be effective and convenient platforms for screening applications, including wound healing and microbial infection of the lip epithelium. Discussion: The establishment of immortalized keratinocytes derived from healthy and cleft lips represents a significant achievement in lip research. These cell lines and the associated three-dimensional lip models are valuable tools that can be used as convenient screening platforms for various assays in a multitude of lip-related research areas, including dermatology, skin care, wound healing, tissue engineering, and craniofacial anomalies. This work opens new avenues in studying lip abnormalities and provides unique tools for personalized medicine approaches beneficial to patients. [ABSTRACT FROM AUTHOR]

Objectives: Cell models have shown great promise as tools for research, potentially providing intriguing alternatives to animal models. However, the original tissue characteristics must be maintained in culture, a fact that is often assumed, but seldom assessed. We aimed to follow the retention of the original tissue identities of cleft lip‐derived skin and mucosa keratinocytes in vitro. Methods: Cleft lip‐derived keratinocytes were isolated from discarded tissue along the cleft margins during cheiloplasty. Cell identities were assessed by immunohistochemistry and quantitative real‐time PCR for tissue‐specific markers and compared with native lip tissue. Moreover, keratinocytes were regularly analyzed for the retention of the original tissue characteristics by the aforementioned methods as well as by differentiation assays. Results: The various anatomical zones of the human lip could be distinguished using a panel of differentiation and functional‐based markers. Using these markers, retention of the original tissue identities could be followed and confirmed in the corresponding primary keratinocytes in culture. Conclusions: Our findings promote patient‐derived cells retaining their original identities as astonishing and clinically relevant in vitro tools. Such cells allow a better molecular understanding of various lip‐associated pathologies as well as their modeling in vitro, including but not restricted to orofacial clefts. [ABSTRACT FROM AUTHOR]

Objectives: To describe the step-by-step procedure of a novel surgical technique consisting of a combination of the laterally closed tunnel (LCT) and the modified coronally advanced tunnel (MCAT) (ie, LCT/MCAT), designed to treat multiple mandibular adjacent gingival recessions (MAGR) and to present the clinical outcomes obtained in 11 consecutively treated patients. Method and materials: Eleven systemically and periodontally healthy patients (7 females, mean ± SD 33.62 ± 14.6 years, min. 19 years max. 67 years) with a total of 40 adjacent mandibular RT1 (ie, Miller Class 1 and 2) gingival recessions with a minimum depth = 3mm, were consecutively treated with LCT/MCAT, in conjunction with an enamel matrix derivative (EMD) and subepithelial palatal connective tissue graft (SCTG). Treatment outcomes were assessed at baseline and at 12 months postoperatively. Prior to surgery and at 12months postoperatively, recession depth (RD) and recession width (RW) were evaluated. The primary outcome variable was complete root coverage (CRC, ie 100% root coverage), the secondary outcome was mean root coverage (MRC). Results: Postoperative pain and discomfort were low and the healing was uneventful in all cases without any complications. At 12 months, statistically significant (P < .05) root coverage (RC) was obtained in all patients. CRC was obtained in five patients with a total of 21 recessions, while MRC measured 92.9% (ie, 3.75 mm). In seven patients (ie, 63.6%), RC amounted to > 93% while the minimum RC per patient measured 83.76%. Conclusion: The results of the present case series suggest that the LCT/MCAT is a valuable technique for the treatment of mandibularRT1 MAGR. [ABSTRACT FROM AUTHOR]

Objective: Transgingival probing is often used in the clinic to assess gingival thickness. However, what is not completely known is how well this method represents the true value of soft tissue thickness. The aim of this study was to assess differences and variation in gingival thickness when measured with transgingival probing or scanned with an intraoral device. Method and materials: This ex vivo study evaluated gingival thickness on 20 porcine cadavers. Gingival thickness was assessed at both central and lateral mandibular incisors through transgingival probing with a standard metal periodontal probe and also using intraoral scanning, which was considered as the method providing the 'true value' of soft tissue thickness. Intra-examiner repeatability and method error were evaluated. Results: No evidence of systematic difference for any of the mandibular central or lateral incisors (mandibular right incisors: mean difference -0.17 to -0.01 mm, and mandibular left incisors: mean difference -0.11 to 0.04 mm) was observed between the periodontal probe and intraoral scanning methods. The absolute differences between the repeated measurements with intraoral scanning for each tooth type (n = 30) were calculated: the overall median was 0.089 mm and the interquartile range was 0.080 mm. Conclusions: Transgingival probing with a standard metal periodontal probe for assessing gingival thickness is a reliable method, with values very close to the true gingival thickness, and it can thus be considered as the clinical gold standard. [ABSTRACT FROM AUTHOR]

Purpose: To use and evaluate two methods for measuring gingival thickness (GT) at mandibular incisors of orthodontic patients and compare their performance in assessing periodontal anatomy through soft tissue thickness.Materials and Methods: The sample consisted of 40 consecutive adult orthodontic patients. GT was measured just before bracket placement at both central mandibular incisors, mid-facially on the buccal aspect, 2 mm apically to the free gingival margin with two methods: clinically with an ultrasound device (USD) and radiographically with cone-beam computed tomography (CBCT).Results: CBCT measurements were consistently higher than USD measurements, with the difference ranging from 0.13 mm to 0.21 mm. No statistically significant difference was noted between the repeated CBCT measurements at the right central incisor (bias = 0.05 mm; 95% CI = -0.01, 0.11; p = 0.104). Although the respective results for the left incisor statistically indicated that the measurements were not exactly replicated, the magnitude of the point estimate was small and not clinically significant (bias = 0.06 mm; 95% CI = 0.01, 0.11; p = 0.014). Small differences between CBCT measurements made by the 2 examiners at the left central incisor (bias = 0.06 mm; 95% CI = 0.01, 0.11; p = 0.014) were detected. However, this difference was minor and also not clinically significant. The respective analysis on the right incisor showed no statistically significant difference (bias = 0.05 mm; 95% CI = -0.01, 0.11; p = 0.246).Conclusions: Based on reproducibility, CBCT imaging for gingival thickness assessment proved to be as reliable as ultrasound determination. However, CBCT consistently yielded higher values, albeit at a marginal level, than did the ultrasound device. [ABSTRACT FROM AUTHOR]

Objectives: To evaluate the healing of multiple adjacent type 1 and 2 gingival recessions (RT1 and RT2) treated with the modified coronally advanced tunnel (MCAT) or the laterally closed tunnel (LCT) in conjunction with a cross-linked hyaluronic acid and subepithelial palatal connective tissue grafts. Method and materials: Fifteen healthy patients exhibiting multiple adjacent mandibular or maxillary RT1 and RT2 of a depth of = 2mm, were treated with the MCAT or LCT in conjunction with cross-linked hyaluronic acid and subepithelial palatal connective tissue grafts. Results were assessed at baseline and after a minimum of 6 months. The primary outcome variable was root coverage. Esthetic outcomes were evaluated on photographs using the root coverage esthetic score. Results: Postoperative pain and discomfort were low and no complications occurred. Data analyses were performed at patient level. After a mean follow-up of 17 ± 5.4 months, statistically significant root coverage was obtained in all 15 cases (P < .0001). Complete root coverage was obtained in 3 out of 15 cases (20%). Root coverage amounted to > 95% in three patients, was between 90% and 95% in four patients, and reached 87.5% in another patient. In three further patients root coverage measured 75%, 77%, and 64.6%, respectively. Mean root coverage measured 85.1 ± 23.2%. Mean keratinized tissue width increased from 2.5 ± 1.0 mm to 3.7 ± 0.7 mm (P < .0001) from baseline to follow-up, while mean probing depth showed no statistically significant changes (1.3 ± 0.5 mm vs 1.5 ± 0.5 mm). The mean root coverage esthetic score was 7.9 ± 1.9, while in the three cases exhibiting complete root coverage, a maximum root coverage esthetic score (10) was given for all treated teeth. Conclusion: Within their limits, the present results indicate that the described treatment approach may lead to predictable root coverage of multiple mandibular and maxillary RT1 and RT2. [ABSTRACT FROM AUTHOR]

To clinically evaluate the healing of mandibular Miller Class I and II isolated gingival recessions treated with the modified coronally advanced tunnel (MCAT) or laterally closed tunnel (LCT) combined with hyaluronic acid (HA) and subepithelial connective tissue graft (SCTG). Method and materials: Twelve healthy patients exhibiting one isolated mandibular Miller Class I or II (Cairo Class 1) gingival recession of a depth of = 3 mm, were consecutively treated with the MCAT or LCT in conjunction with HA and SCTG. Treatment outcomes were assessed at baseline and at least 6 months postoperatively. The primary outcome variable was complete root coverage (CRC). Results: Postoperative pain and discomfort were low and no complications such as postoperative bleeding, allergic reactions, abscesses, or loss of SCTG occurred. After a mean follow- up of 18.9 ± 10 months, statistically significant (P < .0001) root coverage was obtained in all 12 defects. CRC was measured in six out of the 12 cases (50%), four cases showed a root coverage of over 95%, while the remaining two cases reached 80% and 85%. Mean root coverage was 96.09%. Mean keratinized tissue width increased from 1.6 ± 0.8 mm to 4.9 ± 1.3 mm (P < .0001) from baseline to follow-up, while mean probing depth showed no statistically significant changes (1.8 ± 0.9 mm vs 1.3 ± 0.5 mm). Conclusion: Within their limits, the present results indicate that the described treatment approach may lead to predictable root coverage of isolated mandibular Miller Class I and II (Cairo Class 1) gingival recessions. [ABSTRACT FROM AUTHOR]