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Decalcification is crucial in histological processing, particularly for studying mineralized tissues like bone. The choice of decalcification method can significantly impact the quality of histological sections and the preservation of tissue morphology. This study aims to establish a standardized protocol for decalcifying rat calvarial bone using a formic acid-formalin-based decalcification solution. The protocol was systematically optimized and evaluated based on various parameters, including decalcification time, formic acid concentration, and tissue integrity preservation. The decalcification process was evaluated through comprehensive assessments, including gross physical examination, chemical analysis, and radiographic imaging techniques. Our result demonstrated that the 10% formic acid concentration proved most effective for decalcifying rat calvarial bone samples within eight days, excelling in mineral content removal while preserving specimen structural integrity. In contrast, the 5% concentration failed to complete decalcification within ten days, and the 15% compromised sample quality within eight days. Histological analyses confirmed the efficacy of the 10% formic acid concentration in maintaining tissue integrity and achieving optimal staining quality. The standardized protocol presented in this study provides an effective and reliable approach for achieving consistent and high-quality histological sections of rat calvarial bone. An ideal decalcification agent should effectively remove calcium salts, preserve structural integrity and molecular components, facilitate rapid yet minimally damaging decalcification, and ensure ease of handling for laboratory personnel. Further exploration of its applicability to different bone types or species is recommended to broaden its research utility. [ABSTRACT FROM AUTHOR]

Liver cirrhosis poses a global health challenge marked by significant prevalence and mortality. Current therapeutic options are limited by high costs and immune-mediated rejection, necessitating the exploration of innovative strategies to enhance hepatic self-rehabilitation, and counteract the underlying pathological mechanisms. We evaluated the hepatoprotective activity of rat adipose-derived mesenchymal stem cells (ADMSCs) in combination with platelet-rich plasma (PRP) and recombinant human hepatocyte growth factor (rh-HGF) on a rat model of liver fibrosis/cirrhosis induced by bile duct ligation (BDL). Treatment with PRP or rh-HGF alone did not yield significant hepatoprotection in the BDL-induced liver cirrhosis model. However, ADMSC transplantation alone exhibited the potential to alleviate impaired liver conditions. The combination of PRP and rh-HGF demonstrated superior ameliorative effects compared to either treatment alone. Notably, the combination of ADMSC + PRP or ADMSC + rh-HGF significantly enhanced hepatoprotective capacity compared to individual or combined PRP and rh-HGF therapies. Injection of ADMSC via the tail vein reduced inflammation, hepatocyte damage, and collagen deposition, improving overall liver function. This improvement was more pronounced when ADMSC was administered with PRP and rh-HGF versus monotherapy. Our study concludes that ADMSCs exert antifibrotic effects by inhibiting hepatic stellate cell proliferation, collagen synthesis, and inducing apoptosis. ADMSCs also demonstrate immune-modulatory effects and transdifferentiate into hepatic progenitor cells, secreting trophic factors, cytokines, and chemokines that promote impaired liver regeneration. The observed arrest in liver fibrosis progression highlights the potential therapeutic impact of these interventions. [ABSTRACT FROM AUTHOR]

Bone regeneration poses a significant challenge in the field of tissue engineering, prompting ongoing research to explore innovative strategies for effective bone healing. The integration of stem cells and nanomaterial scaffolds has emerged as a promising approach, offering the potential to enhance regenerative outcomes. This study focuses on the application of a stem cell-laden nanomaterial scaffold designed for bone regeneration in rabbits. The in vivo study was conducted on thirty-six healthy skeletally mature New Zealand white rabbits that were randomly allocated into six groups. Group A was considered the control, wherein a 15 mm critical-sized defect was created and left as such without any treatment. In group B, this defect was filled with a polycaprolactone–hydroxyapatite (PCL + HAP) scaffold, whereas in group C, a PCL + HAP-carboxylated multiwalled carbon nanotube (PCL + HAP + MWCNT-COOH) scaffold was used. In group D, a PCL + HAP + MWCNT-COOH scaffold was used with local injection of bone morphogenetic protein-2 (BMP-2) on postoperative days 30, 45, and 60. The rabbit bone marrow-derived mesenchymal stem cells (rBMSCs) were seeded onto the PCL + HAP + MWCNT-COOH scaffold by the centrifugal method. In group E, an rBMSC-seeded PCL + HAP + MWCNT-COOH scaffold was used along with the local injection of rBMSC on postoperative days 7, 14, and 21. For group F, in addition to the treatment given to group E, BMP-2 was administered locally on postoperative days 30, 45, and 60. Gross observations, radiological observation, scanning electron microscopic assessment, and histological evaluation study showed that group F displayed the best healing properties, followed by group E, group D, group C, and B. Group A showed no healing with ends blunting minimal fibrous tissue. Incorporating growth factor BMP-2 in tissue-engineered rBMSC-loaded nanocomposite PCL + HAP + MWCNT-COOH construct can augment the osteoinductive and osteoconductive properties, thereby enhancing the healing in a critical-sized bone defect. This novel stem cell composite could prove worthy in the treatment of non-union and delayed union fractures in the near future. [ABSTRACT FROM AUTHOR]

Wound healing and skin regeneration involve intricate interactions between various cellular, molecular, and biochemical factors. This narrative review aims to provide an in-depth analysis of the present status of therapeutic strategies for wound healing and skin regeneration. The literature review was performed using the Google Scholar search engine with the help of relevant keywords. Selected publications were used to synthesize different sections of the narrative review. The quest for innovative therapeutic approaches to accelerate wound healing and enhance skin regeneration has led to remarkable advancements in recent years. The landscape of therapeutic approaches for wound healing and skin regeneration is evolving rapidly, driven by groundbreaking discoveries and interdisciplinary collaborations. From advanced wound dressings and growth factor therapies to stem cell-based interventions and gene editing techniques, the arsenal of tools at our disposal continues to expand. As researchers continue to unravel the intricate mechanisms underlying wound repair and regeneration, the potential for transformative therapies to revolutionize patient care remains immense. Through a combination of innovative technologies, personalized approaches, ethical considerations, and global accessibility, the future of wound healing holds promise for improving the lives of countless individuals worldwide. Despite significant advancements, several knowledge gaps persist in the field of wound healing and skin regeneration. Further elucidation of cellular and molecular mechanisms governing wound repair, inflammation resolution, and scar formation is warranted. Exploring the crosstalk between wound healing and the microbiome and the influence of ageing and systemic diseases will unravel new therapeutic targets and strategies. As researchers delve deeper into understanding the intricate mechanisms underlying wound repair, the development of novel therapies and their clinical translation become increasingly promising. With a multidisciplinary approach and ongoing advancements in technology, biology, and medicine, the future holds great potential for transforming the field of wound healing and skin regeneration. [ABSTRACT FROM AUTHOR]

Background/Aims: All body functions are activated, synchronized and controlled by a substantial, complex network, the nervous system. Upon injury, pathophysiology of the nerve injury proceeds through different paths. The axon may undergo a degenerative retraction from the site of injury for a short distance unless the injury is near to the cell body, in which case it continues to the soma and undergoes retrograde neuronal degeneration. Otherwise, the distal section suffers from Wallerian degeneration, which is marked by axonal swelling, spheroids, and cytoskeleton degeneration. The objective of the study was to evaluate the potential of mesenchymal stem cell laden neural scaffold and insulin-like growth factor I (IGF-I) in nerve regeneration following sciatic nerve injury in a rat model. Methods: The animals were anaesthetized and a cranio-lateral incision over left thigh was made. Sciatic nerve was exposed and crush injury was introduced for 90 seconds using haemostat at second locking position. The muscle and skin were sutured in routine fashion and thus the rat model of sciatic crush injury was prepared. The animal models were equally distributed into 5 different groups namely A, B, C, D and E and treated with phosphate buffer saline (PBS), carbon nanotubes based neural scaffold only, scaffold with IGF-I, stem cell laden scaffold and stem cell laden scaffold with IGF-I respectively. In vitro scaffold testing was performed. The nerve regeneration was assessed based on physico-neuronal, biochemical, histopathological examination, and relative expression of NRP-1, NRP-2 and GAP-43 and scanning electron microscopy. Results: Sciatic nerve injury model with crush injury produced for 90 seconds was standardized and successfully used in this study. All the biochemical parameters were in normal range in all the groups indicating no scaffold related changes. Physico-neuronal, histopathological, relative gene expression and scanning electron microscopy observations revealed appreciable nerve regeneration in groups E and D, followed by C and B. Restricted to no regeneration was observed in group A. Conclusion: Carbon nanotubes based scaffold provided electro-conductivity for proper neuronal regeneration while rat bone marrow-derived mesenchymal stem cells were found to induce axonal sprouting, cellular transformation; whereas IGF-I induced stem cell differentiation, myelin synthesis, angiogenesis and muscle differentiation. [ABSTRACT FROM AUTHOR]

Twenty clinically healthy adult New Zealand white rabbits of either sex were randomly divided into three groups (I, II and III). Group I and II consisted of eight animals each and in the control group (III) having four animals. Under ketamine (44mg/kg, IM) and xylazine (5 mg/kg, IM) anaesthesia a full thickness abdominal wall defect (2x3 cm) was created and immediately repaired with 0.5% glutaraldehyde treated bovine diaphragm in groups I and II. In control group the defect was repaired by autograft. In group II ultrasound therapy was given at the repair site from day 3 postoperative at the rate of 0.5 watt/cm2 for 5 minutes for two weeks on alternate day. The rectal temperature remained significantly higher in all the groups for 3 days. The swelling, exudation, warmth and pain were maximum at day 1 and then became normal by 6-7 days. Significant rise in neutrophils with reduction in lymphocytes was seen for 7 days in all groups. The grafts were very well taken up by host without any immune reaction. Ultrasound therapy reduced the adhesions in group II. Hexosamine level reduced in all the groups at day 21. Collagen, hydroxyproline and elastin increased at day 21. Changes in zinc and copper was statistically non-significant. Rise in glucose and alkaline phosphatase were although significant but transient. Normal inflammatory cellular reaction was seen in all groups. The healing was seen without any complication in all groups. [ABSTRACT FROM AUTHOR]