Delivery of affordable and scalable encapsulated allogenic/autologous mesenchymal stem cells in coagulated platelet poor plasma for dental pulp regeneration

The main goal of regenerative endodontics procedures (REPs) is to revitalize teeth by the regeneration of healthy dental pulp. In this study, we evaluated the potential of combining a natural and accessible biomaterial based on Platelet Poor Plasma (PPP) as a support for dental pulp stem cells (DPSC) and umbilical cord mesenchymal stem cells (UC-MSC). A comparison study between the two cell sources revealed compatibility with the PPP based scaffold with differences noted in the proliferation and angiogenic properties in vitro. Additionally, the release of growth factors including VEGF, HGF and DMP-1, was detected in the media of cultured PPP and was enhanced by the presence of the encapsulated MSCs. Dentin-Discs from human molars were filled with PPP alone or with MSCs and implanted subcutaneously for 4 weeks in mice. Histological analysis of the MSC-PPP implants revealed a newly formed dentin-like structure evidenced by the expression of Dentin sialophosphoprotein (DSPP). Finally, DPSC induced more vessel formation around the dental discs. This study provides evidence of a cost-effective, xenofree scaffold that is compatible with either autologous or allogenic strategy for dental pulp regeneration. This attempt if successfully implemented, could make REPs treatment widely accessible, contributing in improving global health conditions.

Preparation Technique Affects Recipient Immune Targeting of Autologous Mesenchymal Stem Cells

Fetal bovine serum (FBS) is used for MSC preparation in pre-clinical animal models and veterinary applications, recently in US clinical trials, and for MSC products with current foreign market authorizations. The effect of anti-bovine titers, which are common in animals and humans, has not been investigated. In the equine model, where anti-bovine titers are universally high due to routine vaccination, we evaluated the recipient immune response to autologous MSCs prepared with and without FBS. Preparation of MSCs with FBS resulted in post injection inflammation and antibody mediated cytotoxicity of MSCs when compared to MSCs prepared without FBS. Importantly, synovial MSC concentrations were reduced and LPS induced pain was higher, when FBS was used to prepare MSCs, demonstrating reduced efficacy of FBS prepared MSCs. Fetal bovine serum should no longer be utilized for MSC preparation in pre-clinical study, clinical study, or veterinary applications. The use of FBS in previously reported studies, and in MSC therapeutics with current foreign market authorization, should be considered when interpreting results.

The role of TGF-β2 in cartilage development and diseases

Transforming growth factor-beta2 (TGF-β2) is recognized as a versatile cytokine that plays a vital role in regulation of joint development, homeostasis, and diseases, but its role as a biological mechanism is understood far less than that of its counterpart, TGF-β1. Cartilage as a load-resisting structure in vertebrates however displays a fragile performance when any tissue disturbance occurs, due to its lack of blood vessels, nerves, and lymphatics. Recent reports have indicated that TGF-β2 is involved in the physiological processes of chondrocytes such as proliferation, differentiation, migration, and apoptosis, and the pathological progress of cartilage such as osteoarthritis (OA) and rheumatoid arthritis (RA). TGF-β2 also shows its potent capacity in the repair of cartilage defects by recruiting autologous mesenchymal stem cells and promoting secretion of other growth factor clusters. In addition, some pioneering studies have already considered it as a potential target in the treatment of OA and RA. This article aims to summarize the current progress of TGF-β2 in cartilage development and diseases, which might provide new cues for remodelling of cartilage defect and intervention of cartilage diseases.

Indirect revascularization in patients, suffering critical ischemia of the lower extremities on background of diabetes mellitus

Objective. Determination of clinical efficacy of the mesenchymal stem cells transplantation in patients, suffering critical ischemia of the lower extremities on background of diabetes mellitus. Materials and methods. There were examined 11 patients, suffering critical ischemia of the lower extremities on background of diabetes mellitus Type II with purulent-necrotic processes of the foot, in whom conditions for the direct revascularization of extremity were absent. Microbiological monitoring for the wound discharge, determination of the ankle brachial index and transcutaneous oximetry was performed. The technology elaborated have consisted of two stages. On the first of them a purulent focus sanation (in all the patients the distal amputations of the foot, preserving arterial arch) was done. On the second stage the therapy of the wounds was conducted, using negative pressure, injections of cellular suspension (mesenchymal stem cells) while applying long fine cannula under aponeurosis into the muscles along obliterated vessels in the (10 ± 5) ml quantity with subsequent closure of the wound surface, using fibroblastic matrix. Results. In a year after the treatment critical ischemia in 1 patient was noted, the ischemia Grade III - in 3, and in 7 a IIB Grade have persisted. Conclusion. Performance of indirect revascularization, using autologous mesenchymal stem cells, constitutes an effective and secure procedure.

Autologous Minimally Invasive Cell-Based Therapy for Meniscal and Anterior Cruciate Ligament Regeneration

The meniscus is a fibrocartilaginous tissue that acts as a “shock absorber,” along with performing functions such as stabilization and lubrication of the joint, proprioception, and load distribution. Sudden twisting movements during weight bearing or trauma can cause injury to the menisci, which leads to symptoms such as pain, swelling, and difficulty in performing movements, among others. Conventional pharmacological and surgical treatments are effective in treating the condition; however, do not result in regeneration of healthy tissues. In this report, we highlight the role of cell-based therapy in the management of medial and lateral meniscal and anterior cruciate ligament tears in a patient who was unwilling to undergo surgical treatment. We injected autologous mesenchymal stem cells obtained from the bone marrow and adipose tissue and platelet-rich plasma into the joint of the patient at the area of injury, as well as intravenously. The results of our study corroborate with those previously reported in the literature regarding the improvement in clinical parameters and regeneration of meniscal tissue and ligament. Thus, based on previous literature and improvements noticed in our patient, cell-based therapy can be considered a safe and effective therapeutic modality in the treatment of meniscal tears and cruciate ligament injury.

Evaluation of the Possibility of Fixation of Autologous Mesenchymal Stem Cells in Liver Tissue Introduced into the Portal Vein of the Liver in a Patient Suffering from Cirrhosis

To date, there are quite a large number of reports of the use of stem cells in the treatment of various diseases in humans. However, the authors do not provide credible evidence that the introduced stem cells come directly into the target organ. Without a clear understanding of the location of the introduced stem cells, it is difficult to reliably assess the effect of their use. The article gives its own observation about the possibility of imaging in the human body autologous mesenchymal stem cells introduced through the portal vein of the liver to a patient suffering from cirrhosis. Iron oxide nanoparticles were used as labels. A rather simple question was previously solved: how to create a label of such size that the viability of the stem cell was preserved and it could be visualized in the human body when performing magnetic resonance imaging of abdominal organs. Following in vitro studies, the most appropriate stem cell marking embodiment was chosen to provide both adequate imaging and cell viability. After administration of marked table cells, magnetic resonance imaging of abdominal organs was performed to the patient. The introduced stem cells were visualised only in hepatic tissue. According to clinical and biochemical blood tests - no deviations were detected. There were no complications against the background of this procedure. The patient was discharged for outpatient treatment in satisfactory condition.

Autologous Mesenchymal Stem Cells in The Treatment of Spinal Aneurysmal Bone Cyst

Purpose: We retrospectively analysed a cohort of patients treated at our Centre with bone marrow concentrated (BMC) injection for aneurysmal bone cyst (ABC) of the spine, in order to propose this treatment as a valid alternative for the management of ABCs.Methods: Fourteen patients (6 male, 8 female) were treated between June 2014 to December 2019 with BMC injection for ABC of the spine. The mean age was 17.85 years. The mean follow up was 37.4 months (range 12- 60 months). The dimension of the cyst and the degree of ossification were measured by Computed Tomography (CT) scans before the treatment and during follow-up visits. Results: Six patients received a single dose of BMC, five patients received two doses and in three patients three doses of BMC were administered. The mean ossification of the cyst (expressed in Hounsfield units) increased statistically from 43.48±2.36 HU to 161.71±23.48 HU during follow-up time and the ossification was associated to an improvement of the clinical outcomes. The mean ossification over time was significantly higher in patients treated with a single injection compared to patients treated with multiple injections. No significant difference in ossification was found between cervical and non-cervical localization of the cyst. Moreover, the initial size of the cyst was not statistically associated with the degree of ossification during follow-up Conclusions: Results of this paper reinforce our previous evidence on the use of BMC as a valid alternative for spinal ABC management when SAE treatment is contraindicated or ineffective.