Recellularization of Acellular Xeno Kidney Scaffold: An In Vivo Method to Generate Bioartificial Kidney.

A significant hurdle for kidney tissue engineering is reproducing the complex three-dimensional structure of the kidney. In our study, a stepwise approach of generating a reproducible Xeno kidney scaffold from a goat kidney is described, which can be implanted and recellularized by host cells. We have proposed a combination of sodium dodecyl sulfate and Triton-X-100-based protocol to generate a reproducible Xeno kidney scaffold, which was then analyzed by histology, DNA quantification, SEM, and renal angiography. Further, a small portion from the cortico-medullar region of the acellular scaffold was implanted in the rat's kidney subcapsular pocket for a period of 1 month, to check the recruitment of host cells into the scaffold. Post implantation, the extracellular matrix of the scaffold was well preserved and it did not induce any damage or inflammation in the native kidney. Implantation of the Xeno scaffold resulted in apparent early vascularization which helped in the recruitment of the host cells, which was characterized by histology, immunohistochemistry, and scanning electron microscopy. Implanted Xeno scaffold showed AQP-1, Nephrin, α-SMA, and VEGF expression in proximal tubules and renal glomerulus. Importantly, Ki-67 and WTAP-expressing cells were also observed near proximal tubules suggesting a high level of proliferation in the scaffold. Thus, showing the potential of Xeno kidney development that can be recellularized by the host cell to engineer into a functional kidney.

Silver-doped hydroxyapatite laden chitosan-gelatin nanocomposite scaffolds for bone tissue engineering: an in-vitro and in-ovo evaluation.

Despite the advancements in bone tissue engineering, the majority of implant failures are caused due to microbial contamination. So, efforts are being made to develop biomaterial with antimicrobial property enhancing the regeneration of damaged bone tissue. In the present study, chitosan-gelatin (CG) scaffolds containing silver-doped hydroxyapatite (AgHAP) nanoparticles at 0.5%, 1.0% and 1.5% (w/v) were fabricated by lyophilization technique. The results confirmed the synthesis of AgHAP nanoparticles and showed interconnected porous structure of the nanocomposite scaffolds with 89%-75% porosity. Similarly, the swelling percentage, degradation behavior and compressive modulus of CG-AgHAP nanocomposite scaffolds were 1666%, 40% and 0.7 MPa, respectively. The developed nanocomposite scaffolds revealed better antimicrobial properties and bioactivity. The cell culture studies showed favorable viability of Wharton's jelly stem cells on CG-AgHAP nanocomposite scaffolds. CAM (chorioallantoic membrane) assay determined the angiogenic potential with better visualization of blood vessels in the CAM area. Hence, the obtained results confirmed that CG-AgHAP3 nanocomposite scaffold was the most suitable for bone tissue engineering applications among all scaffolds.

A biologically functional bioink based on extracellular matrix derived collagen for 3D printing of skin.

Over the past few years, several advancements have been made to develop artificial skin that mimics human skin. Artificial skin manufactured using 3D printing technology that includes all epidermal and dermal components, such as collagen, may offer a viable solution. The skin-specific bioink was derived from digested chicken skin and incorporated into PVA (polyvinyl alcohol) and gelatin. The prepared bioink was further analyzed for its structure, stability, biocompatibility, and wound healing potential in in vitro, in ovo, and in vivo models. The 3D-printed skin showed excellent mechanical properties. In vitro scratch assays showed the proliferation and migration of cells within 24 h. In an in ovo assay, the 3D-printed skin facilitated the attachment of cells to the scaffolds. In the animal study, the quick cellular recruitment at the injury site accelerated wound healing. Further, hydroxyproline content was estimated to be 0.9-1.2 mg/ml, and collagen content was 7.5 %, which confirmed the epithelization. The relative expressions of MMP-9, COMP, TNF-α, and IL-6 genes were found to be increased compared to the control. These results demonstrate that 3D bioprinting represents a suitable technology to generate bioengineered skin for therapeutic and industrial applications in an automated manner.

Decellularized small intestine scaffolds: a potential xenograft for restoration of intestinal perforation.

Small intestine perforation is a serious medical condition that requires immediate medical attention. The traditional course of treatment entails resection followed by anastomosis; however, it has complications such as small bowel syndrome (SBS), anastomotic leakage, and fistula formation. Here, a novel strategy is demonstrated, that utilizes the xenogeneic, decellularized goat small intestine as a patch for small intestine regeneration in cases of intestinal perforation. The goat small intestine scaffold underwent sodium dodecyl sulfate decellularization, which revealed consistent, quick, and effective decellularization. Decellularization contributed the least amount of extracellular matrix degradation while maintaining the intestinal architecture. By implanting the decellularized goat small intestine scaffolds (DGSIS) on the chorioallantoic membrane (CAM), no discernible loss of angiogenesis was seen in the CAM region, and this enabled the DGSIS to be evaluated for biocompatibility in ovo. The DGSIS was then xeno-transplanted as a patch on a small intestine perforation rat model. After 30 days post transplant, barium salt used as contrast gastrointestinal X-ray imaging revealed no leakage or obstruction in the small intestine. Histology, scanning electron microscopy, and immunohistochemistry assisted in analyzing the engraftment of host cells into the xeno patch. The xeno-patch expressed high levels of E-cadherin, α-smooth muscle actin (α-SMA), Occludin, Zonnula occluden (ZO-1), Ki 67, and Na+/K+-ATPase. The xeno-patch was consequently recellularized and incorporated into the host without causing an inflammatory reaction. As an outcome, decellularized goat small intestine was employed as a xenograft and could be suitable for regeneration of the perforated small intestine.

Activated platelet-rich plasma accelerate endometrial regeneration and improve pregnancy outcomes in murine model of disturbed endometrium.

Platelet Rich Plasma (PRP) contains high concentrations of growth factors, therefore, PRP activation results in their release, stimulating the process of healing and regeneration. The study was conducted to check whether activated platelet-rich plasma (aPRP) treatment can improve regeneration of the endometrium in an experimental model of ethanol-induced disturbed endometrium. Seventy-two female Wistar rats were randomly assigned into the control group, disturbed endometrium (DE) group and aPRP treated group. Activation of PRP was performed by adding thrombin. All the animals were sacrificed on day 1, day 3, day 6 and day 9 and samples were taken from the miduterine horn. Quantification of Cytokine and chemokine profiles of activated and non-activated PRP for CCL2, TNF- α, IL-1ß, CXCL8, CXCL10, IL2, IL4, IL-6 IL-10, IL-12, IL-17A, TGF- ß, IFN-γ was carried out. Functional and structural recovery of the endometrium was analyzed by hematoxylin-eosin (HE) and immunohistochemical (IHC) analyses. HE confirmed proliferated epithelial lining and stromal reconstruction with decreased fibrosis in PRP treated group compared to the DE group. Epithelial thickness in aPRP treated on day 1, day 3, day 6 and day 9 revealed an significant increase (p ≤ 0.05). Significantly stronger IHC expression of alpha smooth muscle actin, Cytokeratin 18, Cytokeratin 19, Connexin-40, E-Cadherin, Claudin-1, Zona Occludin-1was found in the aPRP treated group compared to the DE group. Furthermore, aPRP treatment was associated with birth of live pups. Our results suggest that intrauterine administration of aPRP stimulated and accelerated the regeneration of endometrium in the murine model of disturbed endometrium.

Cerium oxide nanoparticles disseminated chitosan gelatin scaffold for bone tissue engineering applications.

Cell-free and cell-loaded constructs are used to bridge the critical-sized bone defect. Oxidative stress at the site of the bone defects is a major interference that slows bone healing. Recently, there has been an increase in interest in enhancing the properties of three-dimensional scaffolds with free radical scavenging materials. Cerium oxide nanoparticles (CNPs) can scavenge free radicals due to their redox-modulating property. In this study, freeze-drying was used to fabricate CG-CNPs nanocomposite scaffolds using gelatin (G), chitosan (C), and cerium oxide nanoparticles. Physico-chemical, mechanical, and biological characterization of CG-CNPs scaffolds were studied. CG-CNPs scaffolds demonstrated better results in terms of physicochemical, mechanical, and biological properties as compared to CG-scaffold. CG-CNPs scaffolds were cyto-friendly to MC3T3-E1 cells studied by performing in-vitro and in-ovo studies. The scaffold's antimicrobial study revealed high inhibition zones against Gram-positive and Gram-negative bacteria. With 79 % porosity, 45.99 % weight loss, 178.25 kPa compressive modulus, and 1.83 Ca/P ratio, the CG-CNP2 scaffold displays the best characteristics. As a result, the CG-CNP2 scaffolds are highly biocompatible and could be applied to repair bone defects.

Surgical cotton microfibers loaded with proteins and apatite: A potential platform for bone tissue engineering.

Tissue engineering has emerged as the best alternative to replacing damaged tissue/organs. However, the cost of scaffold materials continues to be a significant obstacle; thus, developing inexpensive scaffolds is strongly encouraged. In this study, cellulose microfibers (C), gelatin (G), egg white (EW), and nanohydroxyapatite (nHA) were assembled into a quaternary scaffold using EDC-NHS crosslinking, followed by freeze-drying method. Cellulose microfibers as a scaffold have only received a limited amount of research due to the absence of an intrinsic three-dimensional structure. Gelatin, more likely to interact chemically with collagen, was used to provide a stable structure to the cellulose microfibers. EW was supposed to provide the scaffold with numerous cell attachment sites. nHA was chosen to enhance the scaffold's bone-bonding properties. Physico-chemical, mechanical, and biological characterization of scaffolds were studied. In-vitro using MG-63 cells and in-ovo studies revealed that all scaffolds were biocompatible. The results of the DPPH assay demonstrate the ability of CGEWnHA to reduce free radicals. The CGEWnHA scaffold exhibits the best properties with 56.84 ± 28.45 µm average pore size, 75 ± 1.4 % porosity, 39.23 % weight loss, 109.19 ± 0.98 kPa compressive modulus, and 1.72 Ca/P ratio. As a result, the constructed CGEWnHA scaffold appears to be a viable choice for BTE applications.

Pathophysiology of Spinal Cord Injury and Tissue Engineering Approach for Its Neuronal Regeneration: Current Status and Future Prospects.

A spinal cord injury (SCI) is a very debilitating condition causing loss of sensory and motor function as well as multiple organ failures. Current therapeutic options like surgery and pharmacotherapy show positive results but are incapable of providing a complete cure for chronic SCI symptoms. Tissue engineering, including neuroprotective or growth factors, stem cells, and biomaterial scaffolds, grabs attention because of their potential for regeneration and ability to bridge the gap in the injured spinal cord (SC). Preclinical studies with tissue engineering showed functional recovery and neurorestorative effects. Few clinical trials show the safety and efficacy of the tissue engineering approach. However, more studies should be carried out for potential treatment modalities. In this review, we summarize the pathophysiology of SCI and its current treatment modalities, including surgical, pharmacological, and tissue engineering approaches following SCI in preclinical and clinical phases.

Heparin Immobilization of Tissue Engineered Xenogeneic Small Diameter Arterial Scaffold Improve Endothelialization.

BACKGROUND: Autologous vessels graft (Inner diameter < 6 mm) harvesting always challenged during bypass grafting surgery and its complication shows poor outcome. Tissue engineered vascular graft allow to generate biological graft without any immunogenic complication. The approach presented in this study is to induce graft remodeling through heparin coating in luminal surface of small diameter (Inner diameter < 1 mm) decellularized arterial graft. METHODS: Decellularization of graft was done using SDS, combination of 0.5% sodium dodecyl sulfate and 0.5% sodium deoxycholate and only sodium deoxycholate. Decellularization was confirmed on basis of histology, and DAPI. Characterization of extracellular matrix was analyzed using histology and scanning electron microscopy. Surface modification of decellularized vascular graft was done with heparin coating. Heparin immobilization was evaluated by toluidine blue stain. Heparin-coated graft was transplanted end to end anastomosis in femoral artery in rat. RESULTS: Combination of 0.5% sodium dodecyl sulfate and 0.5% Sodium deoxycholate showed complete removal of xenogeneic cells. The heparin coating on luminal surface showed anti-thrombogenicity and endothelialization. Mechanical testing revealed no significant differences in strain characteristics and modulus between native tissues, decellularized scaffolds and transplanted scaffold. Collectively, this study proposed a heparin-immobilized ECM coating to surface modification offering functionalize biomaterials for developing small-diameter vascular grafts. CONCLUSION: We conclude that xenogeneic decellularized arterial scaffold with heparin surface modification can be fabricated and successfully transplanted small diameter (inner diameter < 1 mm) decellularized arterial graft.

Evaluation of Quality of Life in Type 2 Diabetes Mellitus Patients Using Quality of Life Instrument for Indian Diabetic Patients: A Cross-Sectional Study.

BACKGROUND: Type 2 diabetes mellitus (T2DM) is a chronic metabolic disease with major impact on the quality of life (QoL) in terms of various domains such as social, physical, and mental well-being. AIM: This study aimed to study the factors determining the QoL in T2DM patients. MATERIALS AND METHODS: A prospective, observational study was conducted in a tertiary care hospital for 6 months. Patients of age ≥18 years and diagnosed with T2DM for ≥6 months (with and without comorbidities) were enrolled for the study. The sociodemographic and clinical characteristics were noted in the self-designed pro forma. The QoL was assessed by the Marathi-translated version of QoL Instrument for Indian Diabetes Patients questionnaire of 34 items and 8 domains. The reliability was validated by Cronbach's alpha. The differences were analyzed by Mann-Whitney U-test and Kruskal-Walis test. RESULTS: Out of 153 T2DM patients, majority were elderly males with mean age of 61.23 ± 11.4 years, married (83%), lower-middle income (57%), urban (51.6%), primary education (46.4%), had diabetes for 5 years or less (42.5%), had positive family history of diabetes (32.6%), and were treated by intensive therapy mainly insulin (41.2%). Statistically significant (P < 0.05) association was found between different domains of QoL and family history, hypertension, body mass index, educational status, marital status, income status, treatment, and complications. The domains of diet satisfaction and general health with the least mean estimates of 7.70 ± 2.62 and 8.25 ± 3.08, respectively, were predominantly affected. CONCLUSION: QoL is an important parameter in diabetes treatment modality. Different factors affected QoL in diabetics in our study. Further studies are definitely needed for better data generation at national level.