Autologous Transplantation Of Adipose-derived Stem Cells Research Articles

Green tea epigallocatechin gallate enhances cardiac function restoration through survival signaling expression in diabetes mellitus rats with autologous adipose tissue-derived stem cells.

The present study tests a hypothesis that cardioprotective effects mediated by autologous adipose-derived stem cells (ADSC) in rats afflicted with insulin-dependent diabetes mellitus (IDDM) may be synergistically enhanced by oral treatment with green tea epigallocatechin gallate (EGCG). Wistar rats were divided into sham, DM, DM+ADSC (autologous transplanted 1 × 106 cells per rat), and DM+ADSC+E (E, green tea oral administration EGCG). Heart tissues were isolated from all rats, and investigations were performed after 2-mo treatment. In the sham, DM, and DM+ADSC groups, we found that DM induced cardiac dysfunction (sham and DM) and autologous ADSC transplantation could partially recover cardiac functions (DM and DM+ADSC) in DM rats. Compared with DM+ADSC, significant improvement in cardiac functions can be observed in DM+ADSC+E in echocardiographic data, histological observations, and even cellular protein expression. Oral green tea EGCG administration and autologous ADSC transplantation show synergistically beneficial effects on diabetic cardiac myopathy in DM rats.NEW & NOTEWORTHY Cardiomyopathy can be induced in rats with diabetes mellitus (DM). Heart function can be restored in DM rats with adipose-derived stem cell treatment. Oral epigallocatechin gallate (EGCG) administration synergistically enhances cardiac function in DM rats with stem cell treatment. The EGCG and stem cell treatment cross-effect occurs via survival protein expression.

Intraglandular Transplantation of Adipose-Derived Stem Cells for the Alleviation of Irradiation-Induced Parotid Gland Damage in Miniature Pigs

In a previous study, the authors verified the protective efficacy of adipose-derived stem cells (ADSCs) on the prevention of salivary gland (SG) damage induced by irradiation in mice. As a critical step before implementation in clinical practice, the present study investigated the protective effect of ADSCs in a miniature pig SG model, because miniature pigs share many characteristics with humans. Third-passage autologous ADSCs at a concentration of 4 × 106cells/mL were transplanted by intraglandular injection into parotid glands (PGs) immediately after local irradiation at a single dose of 20Gy. The injection process was repeated twice a week for 6 consecutive weeks. At 12weeks after irradiation, functional and histologic evaluations were performed by measuring salivary flow rate (SFR) and hematoxylin and eosin and periodic acid-Schiff staining. Immunohistochemical andtransmission electron microscopic examinations also were conducted to evaluate amylase (AMY) production, microvessel density (MVD), and microstructural changes. The irradiated PGs showed remarkable decreases in SFR, AMY production, and MVD. However, transplantation of ADSCs alleviated irradiated PG morphology and function by preserving more functional acinar cells and increasing SFR and AMY production. In addition, greater MVD was observed in the ADSC-treated group than in the irradiated group. These results indicated that intraglandular transplantation of autologous ADSCs is an effective method to protect PGs against damage from irradiation in miniature pigs, which might have clinic application in the future.

Human Adipose-Derived Mesenchymal Stem Cells in Cell Therapy: Safety and Feasibility in Different "Hospital Exemption" Clinical Applications

Based on immunomodulatory, osteogenic, and pro-angiogenic properties of adipose-derived stem cells (ASCs), this study aims to assess the safety and efficacy of ASC-derived cell therapies for clinical indications. Two autologous ASC-derived products were proposed to 17 patients who had not experienced any success with conventional therapies: (1) a scaffold-free osteogenic three-dimensional graft for the treatment of bone non-union and (2) a biological dressing for dermal reconstruction of non-healing chronic wounds. Safety was studied using the quality control of the final product (genetic stability, microbiological/mycoplasma/endotoxin contamination) and the in vivo evaluation of adverse events after transplantation. Feasibility was assessed by the ability to reproducibly obtain the final ASC-based product with specific characteristics, the time necessary for graft manufacturing, the capacity to produce enough material to treat the lesion, the surgical handling of the graft, and the ability to manufacture the graft in line with hospital exemption regulations. For 16 patients (one patient did not undergo grafting because of spontaneous bone healing), in-process controls found no microbiological/mycoplasma/endotoxin contamination, no obvious deleterious genomic anomalies, and optimal ASC purity. Each type of graft was reproducibly obtained without significant delay for implantation and surgical handling was always according to the surgical procedure and the implantation site. No serious adverse events were noted for up to 54 months. We demonstrated that autologous ASC transplantation can be considered a safe and feasible therapy tool for extreme clinical indications of ASC properties and physiopathology of disease.

Effects of transplantation of autologous adipose-derived stem cells on osteoporosis in a rabbit ovariectomy model

Objective To observe the effects of transplantation of autologous adipose-derived stem cells (ASCs) on osteoporosis (OP) in a rabbit ovariectomy (OVX) model.Methods A total of fifteen 6-month-old female New Zealand white rabbits were randomly divided into two groups:ovariectomy group (group A,n =12) and sham operation group (group B,n =3).All rabbits were subjected to bilateral ovariectomy in the group A.Six months later,bone mineral density (BMD) of group A and group B were measured by dual energy X-ray absorptiometry (DXA) to check the result of OVX-OP.ASCs harvested from adipose of OP rabbits were cultured to be expanded and differentiated in osteogenic medium in vitro.Osteogenesis was evaluated by alizarin red staining,alkaline phosphatase (ALP) staining and quantitative assays of osteocalcin (OCN).Autologous osteo-induced ASCs were mixed in calcium alginate hydrogel (CAH) and then transplanted in the left distal femurs,while CAH was transplanted in the right distal femurs of OP rabbits.At 12 weeks after implantation,BMD,micro-CT and histomorphological analyses were performed on these rabbits.Results The BMD of femurs in group A rabbits were obviously lower than that of group B rabbits (P ＜ 0.05) at 6 months after OVX.Compared with control group,ASCs cultured in osteo-induction medium had similar proliferation rate as the non-induced cells,but displayed positive ALP and alizarin red staining and OCN contents.At 12 weeks after implantation,the cell-treated femurs displayed higher BMD,bone trabecula number,trabecula thickness and separation than those of control group,while the structure model index and porosity were lower (P ＜ 0.05).Histological examination indicated that the trabecular thickness increased with complete CAH resorption in cell-treated group,while CAH remained in control group.Conclusions Transplantation of autologous ASCs can help strengthen osteoporotic bone in OVX-OP rabbits,providing a novel approach to OP treatment. Key words: Adipose-derived stem cell; Osteoporosis; Ovariectomy ; Rabbit ; Calcium alginate hydrogel

Autologous Transplantation of Adipose-Derived Stem Cells Enhances Skin Graft Survival and Wound Healing in Diabetic Rats

Diabetes can lead to impaired wound healing and skin grafts used surgically for diabetic wounds are often complicated with necrosis, although different therapies have been proposed. Adipose-derived stem cells (ASCs) participate in tissue repair processes and may have a role during impaired wound healing. In this study, autologous transplantation of ASCs was used to determine if it increases angiogenesis and skin graft survival and enhances wound healing in diabetic rats. Adipose-derived stem cells were successfully isolated and cultured. A full-thickness skin graft model was used to determine the effects of locally administered ASCs in 10 rats rendered diabetic (group 1), whereas 10 others served as controls (group 2). Histological examination of skin grafts followed after 1 week. Additionally, immunohistochemical staining intensity of vascular endothelial growth factor (VEGF) and transforming growth factor β3 (TGF-β3) was assessed in all grafts. The gross and histological results showed significantly increased survival, angiogenesis, and epithelialization. Mean area of graft necrosis was significantly less in group 1 than in group 2 (7.49% vs 39.67%, P < 0.001). Statistically significant increase of capillary density, collagen intensity, VEGF, and TGF-β3 expression was noted in group 1 compared with group 2. These findings suggest that autologous ASC transplantation can enhance skin graft survival in diabetic rats through differentiation, vasculogenesis, and secretion of growth factors such as VEGF and TGF-β3. This might represent a novel therapeutic approach in skin graft surgery for diabetic wounds.

Cell Therapy Using Adipose-Derived Stem Cells for Chronic Liver Injury in Mice.

The present study investigated whether transplantation of autologous adipose-derived stem cells (ASCs) administered into the systemic circulation of a mouse with chronic liver injury provides therapeutic efficacy in the absence of any undesirable side effects. The ASCs used were isolated from mice with the same genetic background as the recipient mice and expanded in vitro. For the induction of chronic liver injury, mice were repetitively administered twice a week with CCl4, a well-known hepatotoxin, for a period of 4 weeks. One day after the eighth dose of CCl4, ASC transplantation was performed by tail vein injection and subsequently followed by two additional doses of CCl4 administration. The recipient mice were divided into four groups (vehicle control, 1.5×103, 1.5×104, and 1.5×105 ASCs per mouse). One day after the final CCl4 administration, all mice were sacrificed to assess serum markers and liver histology. The level of serum markers for liver injury and hepatic function did not differ among the four groups. Similarly, no difference was observed in the liver histology between groups. Cell transplantation with ASCs in our model of chronic liver failure did not result in any observable side effects, but from our results, a single application of ASCs seems to be ineffective in improving liver injury.

Adipose-Derived Stem Cells Enhance Bone Regeneration in Vascular Necrosis of the Femoral Head in the Rabbit

Osteonecrosis of the femoral head was induced in rabbits by intramuscular injection of methylprednisolone and vascular occlusion of the capital femoral epiphysis by electrocoagulation. Eight weeks later the animals received no treatment (group A), core decompression by drilling a hole (diameter 1.2 mm) from the outer cortex 2.5 cm distal to the proximal end of the greater trochanter (group B), or injection of 10(7) autologous adipose-derived stem cells (ADSCs) directly into the femoral head (group C). Eight weeks later, microcomputed tomography scans indicated that bone and trabecular volume and density were significantly higher in group C than in other groups. Histology indicated more new bone formation in group C than in other groups. Group C showed strong osteocalcin immunoreactivity in subchondral bone osteoblasts in the necrotic femoral head, whereas few osteocalcin-positive cells were found among osteoblasts in other groups. Thus, autologous ADSC transplantation improved osteogenesis and the microstructure of vascular deprivation-induced osteonecrotic tissue.

Improvement of skin-graft survival after autologous transplantation of adipose-derived stem cells in rats

Skin grafts are frequently used for a variety of indications in plastic and reconstructive surgery. Their necrosis is a common complication, while different therapies have been proposed. Currently, adipose-derived stem cells (ASCs) hold great promise for their angiogenic potential and role during tissue repair. In this study, autologous transplantation of ASCs was used in skin grafts in rats to determine if it increases angiogenesis, skin-graft survival and wound healing. ASCs were isolated, cultured, labelled with fluorescent dye and injected under full-thickness skin grafts in 10 rats (group 1), while 10 others served as controls (group 2). Skin grafts were analysed after 1 week. Collagen's framework was assessed with Masson's trichrome stain and angiogenesis with von Willebrand factor (vWF) immunohistochemistry. In addition, immunohistochemical staining intensity of vascular endothelial growth factor (VEGF) and transforming growth factor b3 (TGFb3) was assessed in all grafts. Mean area of graft necrosis was significantly less in group 1 than in group 2 (6.12% vs. 32.62%, p<0.01). Statistically significant increase of microvessel density, collagen density, VEGF and TGFb3 expression was noted in group 1 compared with group 2 (all: p<0.01). These findings suggest that autologous ASCs transplantation increases full-thickness skin-graft survival and shows promise for use in skin-graft surgery. This might be both due to in situ differentiation of ASCs into endothelial cells and increased secretion by ASCs of growth factors, such as VEGF and TGFb3 that enhance angiogenesis and wound healing.

Transplantation of patient-derived adipose stem cells in YAC128 Huntington's disease transgenic mice

Huntington’s disease (HD) is a genetic neurodegenerative disorder caused by abnormal expansion of CAG in the huntingtin gene. In R6/2 HD transgenic mice, human adipose-derived stem cells (ASCs) can slow disease progression via secretion of multiple paracrine growth factors. In order to prompt autologous ASCs transplantation in HD, we isolated ASCs from subcutaneous adipose tissues from a HD patient and a normal volunteer. ASCs were grown in two different types of stem cell culture media, EGM-2MV (endothelial growth medium-2 MV) or mesenchymal culture medium (MesenPRO). Cell-surface markers CD13, CD29, CD31, CD34, and CD44 were characterized by flow cytometry. BDNF, HGF, IGF, LIF, NGF, and VEGF expressions were determined by RT-PCR. Cell surface markers for HD ASCs were similar to those for normal ASCs. HD ASCs expressed multiple growth factors, and were similar to normal ASCs, except for NGF; however, they can be altered by culture medium. ASCs were transplanted in bilateral striata of 8 month-old YAC128 mice. At 12 months of age, normal ASCs reduced striatal atrophy, while HD ASCs failed to prevent it. Compared to the control YAC128 group, no improvement in Rotarod performance was observed in any of the transplanted YAC128 mice. However, when normal ASCs were transplanted at 12 months, Rotarod performance was maintained for 4 weeks, with detectable transplanted cells in the striatum and periventricular area. In summary, cultured HD patient-derived ASCs express multiple growth factors with the same cell surface markers as those of normal ASCs in vitro. The efficacy of ASCs transplantation in YAC128 transgenic models can be modified, depending on the time window.

Autologous transplantation of adult adipose derived stem cells into rabbit urethral wall

A study was carried out to evaluate the feasibility of autologous adipose derived stem cells (ADSC) transplantation into female rabbits' urethra walls as an alternative to intrinsic urethral regeneration. Inguinal fat pad of 12 New Zealand adult female rabbits were harvested and processed to obtain stromal vascular fraction (SVF). The SVF were platted to isolate ADSC. Before urethral injection, cells were labeled with DiI marker. The urethra wall was injected with 1 x 10(7) autologous cells or saline (sham). The urethra was harvested at 2, 4, and 8 weeks to identify DiI-labeled cells. At 2 and 4 weeks, the ADSCs create a nodule localized in the urethral sub-mucosa. At 8 weeks, the ADSCs spread and integrated with the urethra wall from the initial injection site. This is the first study to demonstrate a successful autologous ADSCs transplantation. It confirms that ADSCs can survive and integrate within the urethral wall.

The role of stem cells in suppurative environments

Purpose: The management of suppurative perianal lesions presents an extremely challenging problem. Stem cells (SC) extracted from certain tissues, such as adipose tissue, can differentiate into various cell types. Therefore, we have tried to use such cells to stimulate healing in a purulent environment. Methods: In the beginning, we designed a phase I clinical trial, involving five patients with Crohn's disease. We inoculated nine fistulas in four patients with autologous adipose‐derived stem cells (ADSC) and were followed at least 8 weeks. Seventy‐five percent became healed, and 25% showed a decrease in output flow. No adverse effects were observed in any patient. This study evidenced that such cells are safe. Then, we started a research line using SC in different suppurative environments. During the course of these studies, we had the opportunity to treat a patient with perianal hidradenitis suppurativa using our current protocol of ADSC transplantation. Eight weeks after injection, patient had no perianal suppuration, and a year later remains well. Discussion: The biological mechanism that underlies the therapeutic success of ADSC transplantation is unknown. Cell differentiation, secretion of growth factors or immunomodulatory effects have been suggested. No ethical conflicts were identified by our Ethics Committee, because the cells were autologous. Conclusions: Our study shows that ADSC are safe for the treatment of suppurative processes. The actual number of patients included and the uncontrolled nature of these pilot studies do not allow demonstration of the effectiveness of the treatment. However, the results encourage the performance of further studies.