Comparative Evaluation of Three Different Demineralisation Protocols on the Physicochemical Properties and Biocompatibility of Decellularised Extracellular Matrix for Bone Tissue Engineering Applications.

Background With three-dimensional (3D) bioprinting emerging as the ultimate pinnacle of personalised treatment for achieving predictable regenerative outcomes, the search for tissue-specific bioinks is on. Decellularised extracellular matrix (DECM), which provides the inherent biomimetic cues, has gained considerable attention. The objective of the present study was to compare the efficacy of three different demineralisation protocols to obtain DECM for bone tissue engineering applications.  Methodology Goat femurs were treated using three demineralisation protocols to obtain DECM. Group A was treated with demineralisation solution at 40 rpm for 14 days, Group B with freeze-thaw cycles and 0.05M hydrochloric acid (HCl) and 2.4 mM ethylenediamine tetra-acetic acid (EDTA) at 40 rpm for 60 days, and Group C with 0.1M HCl at 40 rpm for three days. After washing, neutralization, 0.05% trypsin-EDTA treatment for 24 hours, and lyophilisation, DECM was obtained. Assessments included scanning electron microscope (SEM) analysis, energy dispersive X-ray (EDX) analysis, hematoxylin and eosin (H&E) staining, and biocompatibility analysis.  Results On comparative analysis, the protocol followed by Group C revealed good surface properties with patent and well interconnected pores with an average pore size of 218.87µm. Group C also revealed carbon and oxygen as predominant components with trace amounts of calcium, proving adequate demineralisation. Group C further revealed optimal demineralisation and decellularisation under histological analysis while maintaining biocompatibility. DECM obtained in Group C should be further processed for bioprinting applications.  Conclusion The three protocols explored in this study hold potential, with Group C's protocol demonstrating the most promise for DECM-based bioink applications. Further research is needed to evaluate the suitability of the obtained DECM for preparing tissue-specific bioinks for 3D bioprinting.

Subperiosteal Ridge Augmentation Technique Utilizing Sticky Bone: A Case Series.

This case series presents the application of a novel minimally invasive technique for augmenting ridge defects across four edentulous sites, thereby circumventing common postoperative complications associated with conventional ridge augmentation techniques. The case series details three cases encompassing four edentulous sites with class I and III ridge defects. This approach uses a minimally invasive subperiosteal technique for ridge augmentation, followed by a delayed implant placement. Subperiosteal incisions were made mesial to the edentulous sites, and subperiosteal pouches were created using tunneling instruments. Sticky bone (comprising injectable platelet-rich fibrin (IPRF) and xenograft) was applied to the pouches, followed by Vicryl suturing. The cone-beam computed tomography (CBCT) assessed dimensional changes between baseline and 180 days post-ridge augmentation. Subsequently, during implant insertion after 180 days, bone samples were collected, decalcified using 10% formic acid, and sectioned to a thickness of 5 µm. Histological analysis of the bone samples was conducted using a bright field microscope, while histomorphometric analysis was carried out using Image J software. The modified subperiosteal tunneling technique employed in this case report, coupled with using sticky bone as an augmentation material, demonstrates promise as a reliable method for ridge augmentation.

Efficacy of a Novel Nanohydrogel Formulation Containing Dopamine, Chitosan Nanoparticles, and Tridax procumbens Extract for Enhanced Wound Healing in Human Gingival Fibroblast Cells: An In Vitro Study.

Background Natural compounds and biomaterials, such as nanohydrogels, have gained interest due to their biocompatibility and tissue regeneration potential. A novel nanohydrogel was prepared by employing Tridax procumbens, a traditional plant with anti-inflammatory properties and chitosan nanoparticles and a natural bioadhesive with potent antimicrobial and antioxidant effects and dopamine, which has been shown to regulate angiogenesis and influence cell growth. The objective of this study was to examine how human gingival fibroblast (HGF) cells respond to a nanohydrogel formulation containing dopamine, chitosan nanoparticles, and T. procumbens extract in terms of cell viability and cell migration. Methods From human gingival tissue, fibroblasts were cultured. A nanohydrogel formulation was prepared by combining dopamine, chitosan nanoparticles, and T. procumbens extract. Three groups were evaluated: Group 1 (nanohydrogel containing dopamine, chitosan nanoparticles, and T. procumbens extract (DnCTP)), Group 2 (chitosan nanoparticles and T. procumbens extract (nCTP)), and Group 3(T. procumbens extract (TP)). The MTT assay was used to measure the percentage of cell viability and a scratch assay to observe cell migration in the wounded area at different concentrations. The data were tabulated in Microsoft Excel (Microsoft Corporation, USA) and imported to IBM SPSS Statistics for Windows, version 23.0 (released 2015, IBM Corp., Armonk, NY), and the Mann-Whitney U test was conducted to statistically analyze the cell viability for different concentrations within the three groups. Results The nanohydrogel formulation (DnCTP) showed dose-dependent effects on cell viability with the highest cell viability at 40 µL/mL concentration, and higher concentrations of 80 µL/mL exhibited cytotoxic effects. nCTP and TP showed decreased cell viability at 80 µL/mL concentration (p < 0.05), indicating potential cytotoxicity at higher concentrations. DnCTP showed improved cell migration in the scratch assay as compared to other groups (nCTP and TP), indicating its potential for facilitating wound healing. Conclusion Dopamine, chitosan nanoparticles, and T. procumbens worked together synergistically to create a nanohydrogel formulation (DnCTP) that showed promise for improving wound healing in human gingival fibroblast cells at a dose-dependent concentration, which may therefore work as an excellent wound-healing agent in periodontal and peri-implant therapy.

Three-Dimensional Bioprinting: The Ultimate Pinnacle of Tissue Engineering.

Three-dimensional (3D) bioprinting has emerged as a revolutionary additive manufacturing technology that can potentially enable life-changing medical treatments in regenerative medicine. It applies the principles of tissue engineering for the printing of tissues and organs in a layer-by-layer manner. This review focuses on the various 3D bioprinting technologies currently available, the different biomaterials, cells, and growth factors that can be utilized to develop tissue-specific bioinks, the different venues for applying these technologies, and the challenges this technology faces.

Green Synthesis and the Evaluation of Osteogenic Potential of Novel Europium-Doped-Monetite Calcium Phosphate by Cissus quadrangularis.

Background The quest for an ideal bone grafting material has been ongoing for decades. Calcium phosphate, alone or in combination with other materials in natural bone, has been shown to aid in bone regeneration effectively. Monetite exhibits superior solubility and resorption rates among calcium phosphates, rendering it an optimal choice for bone regeneration applications. However, the degradation rate of the Monetite is much faster than that of all the other calcium phosphates. Hence, we have added Europium onto the matrix to alter the degradation profile and enhance the osteogenic ability of the prepared matrix. Materials and methods An exclusive Europium-Monetite composite was synthesized employing eco-friendly techniques involving Cissus quadrangularis. The osteogenic potential was gauged using the MG-63 cell line through a calcium mineralization assay employing an Alizarin Red solution, collagen estimation, and an alkaline phosphatase (ALP) assay. The composite's cytocompatibility was evaluated using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay across different concentrations ranging from 12.5 µg to 100 µg. Results Scanning electron microscopy (SEM) analysis of the Europium-Monetite composite revealed a sheet-like arrangement in stacks, and the ATR-IR confirmed the presence of elements Ca, P, and Eu. The osteogenic potential, analyzed by ALP activity, calcium mineralization, and collagen staining, was 10% higher than that of the control (Monetite).  Conclusion The prepared novel Europium-Monetite calcium phosphate complex can enhance the osteogenic potential and could be a promising material for bone regeneration/tissue engineering. The newly created Europium-Monetite calcium phosphate complex holds promise for various bone grafting applications, including integration into scaffolds and as a coating for implants.

Development of a Cissus quadrangularis-Doped Extracellular Matrix and a Hyaluronic Acid-Incorporated Scaffold for Periodontal Regeneration: An In Vitro Study.

PURPOSE: The study aimed to analyze whether adding Cissus quadrangularis (CQ) extract and the extracellular matrix of ovine tendon (TENDON) increases the regenerative potential of mesenchymal stem cells produced in hyaluronic acid (HA) scaffolds for tenogenesis. MATERIALS AND METHODS: Fifty grams of powdered CQ was mixed with 250 mL of ethanol to prepare the extract. Two grams of hyaluronic acid powder was added to 100 mL of distilled water to make the HA solution. The ovine tendon was decellularized using a mixture of 10% phosphate-buffered saline (PBS), sodium dodecyl sulfate (SDS), and Triton-X. The hydrogel samples were prepared by mixing the extracellular matrix of tendon, HA, and CQ, after which they were divided into study groups such as HA, HA + CQ, HA + TENDON, and HA + CQ + TENDON. Scanning electron microscopy (SEM) analysis, swelling analysis, differentiation analysis, compression test, compatibility assay, and tenogenesis assay were later conducted. RESULTS: The morphology of the samples was analyzed using SEM. Low levels of swelling of the hydrogels were observed. Cells were found to be viable and showed good differentiation and tenogenesis. Optimal compression levels were observed, and the properties of the prepared hydrogels were satisfactory. CONCLUSION: The results suggest that the addition of CQ considerably increases the tenogenic potential of the extracellular matrix/HA scaffold. Hence, it can be used as a regenerative material for periodontal tissue regeneration.

Three-Dimensional Collagen Membranes Challenging the Gold Standard in Gingival Recession.

Gingival recession is a commonly encountered chief complaint in a dental office, most commonly presenting as pain or sensitivity on intake of food or sometimes just seen as an esthetic concern which may present in a localized or generalized form. Such interest toward dental esthetics has encouraged clinicians to formulate newer minimally invasive surgical techniques which are short but still deliver predictable long-term benefits in restoring the patient's smile. One such method was evaluated in the present short communication where a commercially available, biodegradable, and volume-stable collagen membrane was used to manage gingival recession instead of donor grafts. The clinical parameters which were assessed after a period of 9 months demonstrated an increased width of attached gingiva and overall tissue thickness after surgical intervention. These findings along with successful mean root coverage of the upper left canine would seem to challenge the results obtained while using similar procedures and donor grafts, the age-old gold standard bio-filler. The dawn of esthetic dentistry is here with more emphasis on how physical appearance bolsters confidence and morale among younger individuals. Among the various complaints concerned with patient`s esthetic, periodontal plastic surgery is one of the more challenging procedures. The real obstacle while performing such procedures is achieving complete or partial root coverage over an avascular bed, whereas multiple variables seem to affect the overall clinical outcome.

Impact of COVID-19 on Renal Function: A Multivariate Analysis of Biochemical and Immunological Markers in Patients.

Introduction There have been tremendous continuous efforts to understand the broad spectrum of disease and its sequelae since the start of the coronavirus disease 2019 (COVID-19) pandemic. Several studies have identified biomarkers that correlate with multiple organ failure in COVID-19 patients. The purpose of our study was to evaluate COVID-19-associated kidney injury. Methods This retrospective cross-sectional study was conducted at the Institute of Biochemistry, Madras Medical College, by reviewing the electronic records of 1,000 reverse transcription-polymerase chain reaction (RT-PCR)-confirmed COVID-19-positive patients admitted at the COVID-19 care center. Data were extracted from the case records of 1,000 RT-PCR-positive patients with different CT chest grades plus comorbid conditions such as type 2 diabetes mellitus (T2DM), systemic hypertension (SHT), coronary artery disease (CAD), chronic obstructive pulmonary disease (COPD), and cerebrovascular accident (CVA) as Group I (n = 500). Group II (n = 500) comprised of COVID-19-positive patients with no comorbid conditions. The data were recorded from all the patients at the time of admission, prior to starting treatment. Patients with comorbid and non-comorbid conditions were compared according to different CT grades. Results COVID-19 patients with different CT grades showed a significant relationship with creatinine, sodium, potassium, C-reactive protein (CRP), ferritin, total protein, and albumin with p-values of 0.04, 0.01, 0.02, 0.000, 0.00, 0.00, and 0.000, respectively, in Group I. In Group II, with various grades of CT changes, the neutrophil-lymphocyte ratio (NLR) and creatinine showed no significance. The sodium, potassium, CRP, ferritin, total protein, and albumin showed low significance with the chest CT grades. Conclusions Our study demonstrated that COVID-19 can cause mild to moderate renal impairment in COVID-19 patients. Multiple factors contributed to this, such as the higher angiotensin-converting enzyme 2 (ACE2) expression on kidney cells, microinflammation, increased blood clotting, and probable direct infection of the kidney. A high NLR, increased inflammatory markers, and altered renal function analytes such as urea, creatinine, sodium, potassium, total protein, and albumin also confirmed this.

An atypical presentation of plasma cell gingivitis with generalized skin lesions.

This article aims to present a report of an atypical clinical presentation of a plasma cell gingivitis (PCG) case with unusual skin lesions. PCG is a rare benign inflammatory condition which can be classified into Type 4 hypersensitivity reaction. It occurs due to reaction to unknown antigen, often flavoring agents or spices found in chewing gums, toothpastes and lozenges. Histologically, the lesion shows dense plasma cells infiltrate in the connective tissue. Early diagnosis of PCG is essential to differentiate from variety of conditions, namely, leukemia, HIV infection, discoid lupus erythematosus, atrophic lichen planus, desquamative gingivitis, or cicatricial pemphigoid which must be differentiated through hematologic and serologic testing. In this article, we will discuss a case of PCG with unusual skin lesions.

Analysis of association between TLR-4 Asp299Gly and Thr399Ile gene polymorphisms and chronic periodontitis in a sample of south Indian population.

BACKGROUND: To analyze the association between TLR-4 Asp299Gly and Thr399Ile gene polymorphisms and chronic periodontitis in a sample of south Indian population. MATERIALS AND METHODS: Genomic DNA was obtained from peripheral blood of 60 patients with chronic periodontitis and 60 periodontally healthy subjects. TLR-4 Asp299Gly and Thr399Ile gene polymorphisms were genotyped by a polymerase chain reaction-restriction fragment length polymorphism method. The data were analyzed by a χ(2)-test and by relative risk estimation. RESULTS: Thr399Ile alleles were found in 4% of chronic periodontitis patients and in 1% of periodontally healthy subjects. The prevalence of a Thr399Ile heterozygote was found to be 5% in the chronic periodontitis group and 1.67% in the periodontally healthy group, respectively. Homozygosity for TLR-4 Thr399Ile was seen in chronic periodontitis patients only, which was 1.67%. The TLR-4 Asp299Gly gene polymorphism was not detected in either chronic periodontitis or periodontally healthy groups. CONCLUSION: There is no significant association between TLR-4 Thr399Ile polymorphism and chronic periodontitis in a sample of south Indian population.