In vitro Cytotoxicity Evaluation of Flowable Hyaluronic Acid-Acellular Stromal Vascular Fraction (HA-aSVF) Mixture for Tissue Engineering Applications.

Background: The stromal vascular fraction (SVF) is an aqueous fraction isolated from the adipose tissue that constitutes different kinds of cells and extracellular matrix components. Hyaluronic acid (HA) is a linear polysaccharide in vertebrate tissues and is considered a potential tissue engineering scaffold due to its biocompatible nature. In this study, we have evaluated the cytotoxicity of xenofree HA in combination with an acellular component of adipose SVF (HA-aSVF) to propose it as a candidate biomaterial for future applications. Materials and Methods: 3-(4,5-dimethyl thiazolyl-2)-2,5-diphenyltetrazolium bromide assay of L-929 cells treated with HA-aSVF was used in our study. Data were normalized to cell control (untreated) and extracts of copper and ultra-high molecular weight polyethylene were used as positive (PC) and negative controls (NC). Results: Fibroblast cells retained the morphology after 24 h of treatment with HA-aSVF mixture and exhibited a similar percentage of cell activity compared to NC. PC showed a positive cytotoxic response as expected. The cells incubated with HA-aSVF showed a linear increase in cell activity indicating proliferation. Conclusion: The mixture of HA and acellular SVF in its flowable form is non-cytotoxic and showed improved cell proliferation. Hence the mixture can be proposed as a biomaterial and can be further explored for specific tissue engineering applications.

Evaluation of Non-Cross Linked Xeno-Free Hyaluronic Acid Solutions as Visco Elastic Biomaterials.

Background: Hyaluronic acid (HA) is a naturally occurring biodegradable, high molecular weight, non-sulfated glycosaminoglycan (GAG) polymer known for its excellent biocompatibility. HA-based products are widely used as viscosupplements, dermal fillers, and ophthalmic lubricants in clinical settings. Although animal and bacterial-derived HA are commonly reported, plant-sourced HA is not frequently reported. In this study, we have evaluated various viscoelastic properties of one such plant-based HA solution and propose them as an alternative to existing animal/bacteria-sourced HA. Materials and Methods: The viscoelastic properties of plant-sourced HA solution of various concentrations (0.1%, 0.5%, 1%, and 2% in PBS) were studied using a rheometer at 37°C. Flow curves, amplitude sweep studies, and frequency sweep studies were performed and compared for all HA solutions. Results: The HA solutions displayed shear-thinning behavior, which is an important characteristic of an injectable biomaterial. The 0.1 and 0.5% HA were found to have viscoelastic properties appropriate for eye lubricants, while 1 and 2% HA solutions showed properties suitable for soft tissue fillers. Frequency sweep studies indicated that all the samples are typically viscoelastic liquids with a loss modulus (G″) higher than the storage modulus (G'). This indicated that the samples needed further processing like crosslinking of HA or using higher molecular weight HA to be suitable as viscosupplements. However, the frequency sweep studies also indicated that these solutions can be used as soft tissue fillers of any type based on the G' and tan δ values. Conclusion: The plant-sourced HA solutions are found to exhibit good shear-thinning properties with viscoelastic properties suitable for eye lubricants and soft tissue fillers. However, to be used as viscosupplements, the viscoelastic properties of HA solutions have to be further modified through non-toxic crosslinking strategies, and hydrophobic derivatives as well as by using high molecular weight HAs.

Utility of Chitra-HASi Granules in Cystic Defects of the Maxillofacial Region: A Pilot Study.

BACKGROUND: Cystic defects that are critical sized or larger require bone replacement strategies. However, due to inherent disadvantages of the various types of grafts, none of the available materials are best suited for these defects. Among the alloplastic materials, hydroxyapatite (HA)-based grafts are the most popular, due to their osteoconductive nature and resemblance to mineral bone. The aim of the study was to assess the utility of the novel material "Chitra-HASi" as a bone substitute in the maxillofacial region. MATERIALS AND METHODS: In a single-arm, prospective study, patients with radicular and dentigerous cysts were included and the minimum defect size was standardized at 20 × 20 mm or above. The Chitra-HASi material was developed by a wet precipitation technique and adopted for use following multiple in vitro and in vivo studies, confirming its safety and biocompatibility profile. All cysts underwent enucleation, followed by peripheral ostectomy and apicectomy of the teeth involved. The HASi graft was packed inside the cystic defect in a granular form and covered with a mucoperiosteal flap. Panoramic radiographs were taken preoperatively and at 3, 6, and 12 months postoperatively. RESULTS: Twenty-three patients were included in the study, of which only 10 patients could be followed up for 12 months after graft placement. The mean preoperative bone density was found to be 14.9% ± 4.97 (standard deviation), whereas the postoperative 3-month, 6-month, and 12-month densities had a mean difference of -11.3%, -22.9%, and -37.3%, respectively, and the differences were statistically significant. Minor complications such as sinus formation (n = 7) and extrusion of granules (n = 4) were noted, which were managed conservatively. Only two patients required graft removal secondary to infection, leading to a persistent sinus tract. CONCLUSION: The results of the study suggest that Chitra-HASi granules show potential as an alternative to other bone substitutes. The addition of silica to the porous HA material offers superior strength characteristics and needs long-term evaluation to assess its stability in large cystic defects.

Dental Pulp Stem Cells in Neuroregeneration.

Neurological diseases and injuries affect the routine life of patients. Current medical and surgical treatment has not improved the quality of life to desired limits. Neural regeneration through stem cells may be ideal choice in current scenario. Dental pulp stem cells (DPSCs), which are isolated from dental pulp, have shown excellent neuroregenerative properties in various animal studies. This review outlines the clinical perspective of DPSCs in neuroregeneration.

Evaluation of Antimicrobial Efficacy of Calcium Hydroxide along with Proton Pump Inhibitor against Enterococcus Faecalis.

AIM: The aim of this study was to evaluate in vitro the efficacy of pantoprazole, a proton pump inhibitor, alone, and in combination with calcium hydroxide against Enterococcus faecalis, a star survivor in endodontic pathology. MATERIALS AND METHODS: Teeth were inoculated with E. faecalis strains MTCC 439 and were divided into groups. Group 1 used saline as the negative control, Group 2 used calcium hydroxide, Group 3 used pantoprazole alone, and Group 4 used calcium hydroxide along with pantoprazole. They were incubated and dentine harvesting was performed. Colonies were counted using a digital counting machine. Data were statistically analyzed. RESULTS: Group 4 was found to be the most effective against the pathogen. DISCUSSION: This study indicates that the association of calcium hydroxide with the proton pump inhibitor pantoprazole can be successfully used as an intracanal medicament against E. faecalis.

Reconstruction of Craniofacial Bone Defects with Autologous Human Bone Marrow Stem Cells and Autogenous Bone Grafts: A Case Report with Review of Literature.

Reconstruction of craniofacial bony defects has always been a challenging task for the surgeons over the years. The science of reconstructing such defects is of at most importance to craniofacial and plastic surgeons due to its relevance in facial aesthetics function as well as prerequisite procedure for continuing other surgical procedures. The main goal of the reconstruction of the craniofacial defects is to reduce the morbidity by restoring the facial form and aesthetics, as well as a good function of the facial structures by achieving a reasonable occlusion and articulation. Although significant improvements have occurred during the last few decades, challenges still exist as to what type of reconstruction to be carried out with regard to techniques and the type and quality of materials of choice to be used. As decades progressed, the advancement in surgical techniques and the variety of reconstruction methods have definitely improved the quality of life. This article reviews the method of bony reconstruction of craniofacial defects using autologous human bone marrow stem cells and autologous bone grafts and its modification, which includes much recent tissue engineering techniques and regenerative medicine, thereby replacing older techniques by biological substitutes, which can restore improve and maintain orofacial function and aesthetics.

A smart tele-cytology point-of-care platform for oral cancer screening.

Early detection of oral cancer necessitates a minimally invasive, tissue-specific diagnostic tool that facilitates screening/surveillance. Brush biopsy, though minimally invasive, demands skilled cyto-pathologist expertise. In this study, we explored the clinical utility/efficacy of a tele-cytology system in combination with Artificial Neural Network (ANN) based risk-stratification model for early detection of oral potentially malignant (OPML)/malignant lesion. A portable, automated tablet-based tele-cytology platform capable of digitization of cytology slides was evaluated for its efficacy in the detection of OPML/malignant lesions (n = 82) in comparison with conventional cytology and histology. Then, an image pre-processing algorithm was established to segregate cells, ANN was trained with images (n = 11,981) and a risk-stratification model developed. The specificity, sensitivity and accuracy of platform/ stratification model were computed, and agreement was examined using Kappa statistics. The tele-cytology platform, Cellscope, showed an overall accuracy of 84-86% with no difference between tele-cytology and conventional cytology in detection of oral lesions (kappa, 0.67-0.72). However, OPML could be detected with low sensitivity (18%) in accordance with the limitations of conventional cytology. The integration of image processing and development of an ANN-based risk stratification model improved the detection sensitivity of malignant lesions (93%) and high grade OPML (73%), thereby increasing the overall accuracy by 30%. Tele-cytology integrated with the risk stratification model, a novel strategy established in this study, can be an invaluable Point-of-Care (PoC) tool for early detection/screening in oral cancer. This study hence establishes the applicability of tele-cytology for accurate, remote diagnosis and use of automated ANN-based analysis in improving its efficacy.

Immunotherapy in Oral Cancer.

Immunotherapy is one of the newer entities which is promising, at least can be very much helpful as an adjuvant therapy. This newer modality of the treatment in the field of cancer treatment may be the fourth pillar supporting surgery, chemotherapy, and radiotherapy. Careful selection of patient is the key for success of immunotherapy, which is based on patient's immunological contexture. This review aimed to present the fundamental aspects of tumor immunity and immunotherapy, focused on oral squamous cell carcinoma.

Platelet-Rich Plasma and Platelet-Rich Fibrin in Periodontal Regeneration: A Review.

Platelet concentrates (PCs; platelet-rich plasma and platelet-rich fibrin) are autologous bioactive substances that have found varied application in medical and dental fields, particularly in oral and maxillofacial surgery, plastic surgery, and sports medicine. The rationale of these technologies is to extract all the elements from patient's own blood sample, which could be used to improve healing by promoting tissue regeneration. PCs have evolved a long way since its introduction in 1954. PCs have been used successfully in periodontics and implant dentistry. However, the preparation protocol, processing time, transfer of concentrates, centrifugation temperature, vibration, etc., being not standardized are various factors for the mixed results reported in the literature. This review intends to discuss evolution of PCs, their preparation techniques, and their clinical and technical aspects and applications.

Comparison of Efficacy of Lycopene and Lycopene-Hyaluronidase Combination in the Treatment of Oral Submucous Fibrosis.

BACKGROUND: Oral submucous fibrosis (OSMF) is a chronic progressively scarring disease of the oral cavity. Lycopene is a powerful antioxidant obtained from tomatoes and has the highest singlet oxygen quenching capacity and a high capacity of quenching other free radicals in vitro among dietary carotenoids. Hyaluronidase is a substance prepared from the testes and semen of mammals that modifies the permeability of connective tissue through the hydrolysis of hyaluronic acid. OBJECTIVE: To evaluate the efficacy of lycopene and lycopene-hyaluronidase combination, and to compare the efficacy of lycopene and lycopene-hyaluronidase combination in the treatment of OSMF. STUDY DESIGN: The study consisted of 45 patients with OSMF divided into three equal groups. Patients in Group A were given Lycored 16 mg daily in two equally divided doses for 3 months. Patients in Group B were given LycoRed along with hyaluronidase intralesional injection of 1500 IU twice weekly for 3 months. Patients in Group C were given placebo capsules. Patients were evaluated after 3 months. The following parameters were recorded: mouth opening, visual inspection, palpatory findings, and burning sensation. RESULTS: There was statistically significant change in mouth opening and burning sensation for lycopene and lycopene-hyaluronidase combination than in the placebo group in the treatment of OSMF, but the lycopene-hyaluronidase combination did not show any statistically significant change when compared with lycopene alone. CONCLUSION: Lycopene appears to be a very promising antioxidant in the management of oral submucous fibrosis, both in clinical and symptomatic improvement.

Estimation of Serum Cortisol Levels in Oral Lichen Planus Patients with Electrochemiluminescence.

OBJECTIVES: The objectives of this study were to compare the serum cortisol level in oral lichen planus (OLP) patients with normal healthy individuals and to compare the serum cortisol level in patients with erosive and nonerosive forms of OLP. MATERIALS AND METHODS: A case-control study involving 60 patients within the age group of 20-50 years was carried out. Group A included 15 subjects with nonerosive lichen planus, without skin involvement; Group B included 15 subjects with erosive lichen planus, without skin; and Group C included 30 subjects with no apparent lesions of the oral mucosa and the skin. From the cases and controls, 7mL venous blood was taken from median cubital vein two times (between 8 and 10 AM, and 4 and 6 PM) by using a 10 mL syringe from which 5 mL was taken for estimation of serum cortisol. RESULTS: The serum cortisol levels of the erosive OLP patients were significantly higher than those of nonerosive OLP patients and controls. There was no significant difference in the serum cortisol levels between nonerosive OLP in comparison with controls.

Ultrastructural Changes of Collagen in Different Histopathological Grades of Oral Submucous Fibrosis.

BACKGROUND AND OBJECTIVES: Oral submucous fibrosis (OSMF) is a potentially malignant disorder and a crippling condition of oral mucosa. It is usually seen in adults with areca nut chewing habit, which is characterized by changes in the connective tissue fibers leading to stiffness of the mucosa and restricted mouth opening. Patients with severe cases have distinct difficulties in chewing, swallowing, and speaking. It predominantly occurs in Indians and other population of the Indian subcontinent with a prevalence of 0.2%-0.5%. The potentiality of malignant transformation associated with OSMF is much higher with a range of 4.5%-7.6%. This article is about the ultrastructural changes pertaining to collagen and with respect to the histopathological grades of OSMF. MATERIALS AND METHODS: A total of 20 biopsy samples from clinically diagnosed OSMF subjects and 5 healthy controls were collected. After obtaining the biopsy, the specimens were divided into two halves, of which one was studied under light microscope and the other half was observed under transmission electron microscope. RESULTS: Of the 20 subjects in the study group, most of the subjects were in the 20-29 years age group with an overall male predilection (19 subjects). Four subjects were in grade 1, nine were in grade 2, and seven were in grade 3. Collagen showed sparse to dense fibrosis and normal to thick collagen bundles, and some subjects showed encroachment of collagen into the blood vessels. CONCLUSION: This study showed definitive changes with respect to collagen in the OSMF samples compared to the controls. The changes were found to be increasing with the progression of the disease to the higher grades. The changes were pertaining to the collagen and were drawing toward an increased fibrosis of the connective tissue, which compresses the blood vessels. Hence, the state of hypoperfusion and subsequent epithelial atrophy can be considered in the progression of disease to the higher grades.

Minor Salivary Gland Changes in Oral Epithelial Dysplasia and Oral Squamous Cell Carcinoma - A Histopathological Study.

INTRODUCTION: The most common etiology for Oral Squamous Cell Carcinoma (OSCC) is tobacco and tobacco related products which cause nuclear damage to the keratinocytes. The chemical carcinogens not only affect the lining of oral epithelium but also affect the lining epithelium of the excretory ducts of the salivary glands. Thus, there is a possibility of epithelial dysplasia of the salivary duct epithelium which may lead to potential malignant transformation. AIM: The study was performed to see the changes in the minor salivary glands and excretory ducts in cases of oral epithelial dysplasia and OSCC. MATERIALS AND METHODS: A total of 278 archival cases of mild, moderate and severe epithelial dysplasia, carcinoma in situ, OSCC including verrucous carcinoma were histopathologically evaluated to observe changes in the excretory ducts and the minor salivary glands. RESULTS: In the study there were 56.5% males and 43.5% females. The age group that was most commonly affected in both the sexes was 50-60 yr old. Buccal mucosa was the most common site of involvement. Ductal changes observed in the excretory duct include simple hyperplasia, metaplastic changes such as mucous, oncocytic & squamous, and infiltration of inflammatory cells and malignant cells. Acinar changes observed were degeneration, squamous metaplasia, myoepithelial cell proliferation and inflammatory cell infiltration. Both the excretory ducts and ducts within the gland showed dysplasia. CONCLUSION: According to observations in our study it is suggested that histopathological interpretation for oral mucosal lesions especially oral epithelial dysplasias and OSCC should also include changes related to salivary gland tissue to provide a better treatment plan and prevent recurrence of the malignant tumours.

Immunohistochemical Detection of p75 Neurotrophin Receptor (p75-NTR) in Follicular and Plexiform Ameloblastoma.

INTRODUCTION: Ameloblastoma holds a unique position among benign tumours by its locally destructive and invasive nature. Recently improvised molecular techniques helped researchers to unravel the myth behind such biologic behaviour. Though interesting findings have been delivered, the rhythmic correlation regarding the exact mechanism still remains lacking. Neurotrophins and their receptor mediated pathways play a crucial role in survival, death and differentiation of many neuroectoderm derived cells. With this background, the study has been aimed to investigate the expression of p75-NTR (Neurotrophin Receptor) in follicular and plexiform ameloblastoma. AIM: To analyze the immunohistochemical expression pattern of p75-NTR in ameloblastoma and to compare the immunohistochemical expression pattern of p75-NTR among the histological types of ameloblastoma, follicular and plexiform patterns. MATERIALS AND METHODS: Total 22 ameloblastomas (12 follicular, 10 plexiform) were immuno-stained with anti-human p75-NTR mouse IgG monoclonal antibody and the pattern of staining is statistically analyzed. RESULTS: Only 11 (10 follicular, 1 plexiform) out of 22 ameloblastomas showed immuno-reactivity to p75-NTR. In ameloblastoma, only the peripheral pre-ameloblast like tall columnar cells showed reactivity whereas the stellate reticulum-like cells were immuno-negative. The staining pattern was membranous in the immuno-reactive cells. The results were studied with the downstream pathways from the literature and a possible mechanism has been proposed. CONCLUSION: The expression pattern of p75-NTR was found to be more in follicular ameloblastoma than plexiform.