Real-time effect of adipose stem cells on fibrogenesis of dermal fibroblast co-stimulated by transforming growth factor-β1 / 上海交通大学学报（医学版）

Objective • To mainly explore the real-time effect of adipose stem cells (ASCs) on the fibrogenesis of dermal fibroblasts co-stimulated by transforming growth factor-β1 (TGF-β1), and further clarify the possible pathway and mechanism of ASCs in regulating wound repair. Methods • By using two different real-time culture systems including Transwell system and contact co-culture system, events associated with fibrogenesis including the changes of fibroblast cell number or expression of collagen types and III detected by immunofluorescence or Western blotting in dermal fibroblasts at 72 h with/without the stimulation of transforming growth factor-β1 (TGF-β1) and/or ASCs were studied. Results • In Transwell system, the cell number of fibroblasts was significantly decreased under the stimulation of ASCs and TGF-β1, compared with TGF-β1 only group (P=0.035). In contact co-culture system, under the stimulation of TGF-β1, the numbers of fluorescence labeling fibroblasts in group with ASCs as basal cells were decreased, compared with group with fibroblast as basal cells (P=0.000). In terms of the collagen expression, in Transwell system, the amounts of collagen secretion from fibroblasts within the upper chamber were increased dramatically when fibroblasts were being co-cultured with ASCs (P=0.000). In contact co-culture system, under the stimulation of TGF-β1, the amounts of collagen secretion in the supernatant of cell culture in the group with ASCs as basal cells were increased, compared with the group with fibroblast as basal cells (P=0.000). Conclusion • ASCs may have an effect on fibrogenesis of dermal fibroblasts co-stimulated by TGF-β1 through a paracrine and direct contact way. It not only increases collagen production and secretion, but also inhibits fibroblasts over-proliferation.

Bone Morphogenetic Protein 2-Conjugated Silica Particles Enhanced Early Osteogenic Differentiation of Adipose Stem Cells on the Polycaprolactone Scaffold

BACKGROUND: Silica particles (SPs) induce cell proliferation and osteogenic differentiation. We reported that SPs in the scaffold induced early stage osteogenic differentiation. METHODS: A polycaprolactone (PCL) scaffold was fabricated with a 10 wt% SPs. The surface of PCL scaffold was coated with a 10 µg/mL collagen solution. Next, the scaffold was conjugated with 2 µM SPs, 2 µg/mL bone morphogenetic protein 2 (BMP2), or 2 µM BMP2-conjugated SPs (BCSPs). Green fluorescent protein-coupled BMP2 was applied to fabricate the scaffold. The fluorescence intensity was analyzed by confocal microscopy. The mRNA levels of the early osteogenic differentiation marker, alkaline phosphatase (ALP), were analyzed by real-time quantitative polymerase chain reaction. Levels of BMP2, RUNX2, ERK1/2, and AKT were assessed by western blotting. RESULTS: ALP mRNA levels were significantly higher in the BCSP-conjugated scaffold than in the other scaffolds. In the early stage of osteogenic differentiation, the protein levels of BMP2, RUNX2, ERK1/2, and AKT in cells were significantly higher in the BCSP-conjugated scaffold than in other scaffolds. Thus, the BCSP composite scaffold induced rapid osteogenic differentiation. CONCLUSION: These results suggest that BCSP composite can be used to promote early stage osteogenic differentiation and show promise as a material for use in scaffolds for bone regeneration.

Adipose Stem Cells with Conditioned Media for Treatment of Acne Vulgaris Scar

This study was to investigate the effect of subcutaneous injection of the adipose stem cells (ASCs) with conditioned media (CM) in the treatment of acne vulgaris scar. We used Adult male New Zealand white rabbit ears as an animal model and induced acne formation by Kignman method. Adipose tissue was isolated and harvested from the scapula of rabbits, and ASCs were cultured and expanded until passage 1. There have four groups in our experiment, include phosphate buffered saline (PBS), ASCs with PBS (ASC + PBS), CM, and ASCs with CM (ASC + CM) group. This solution of 0.6 ml injected to subcutaneous in each group. ASC + PBS and ASC + CM groups were containing ASCs of 5.0 × 106 cells/ml. We analyzed the treatment of 4 groups to scar tissue after 2 and 4 weeks by hematoxylin and eosin stain, immunohistochemistry, and RNA expression level of tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α), and matrix metalloproteinase-2 (MMP-2). Also, the expression of keratin 16 (K16) was detected by western blot analysis. H&E stain showed that infiltration of inflammation cells was significantly reduced at 2 and 4 weeks, as well as re-epithelialization was improved in the ASC + CM group. The ASC + CM gourp was reduced both expression levels of TNF-α, IL-1α, and MMP-2 and K16 protein level. In conclusion, the ASCs with CM has a significant curative effect on acne vulgaris scar, more to the point, the CM has a key role on treatment. It could be applied to a therapeutic approach to regenerate to treat acne vulgaris scar.

Recent Advancements in Autologous Fat Grafting

Autologous fat grafting has been performed for more than one hundred years and there had been a major refinement of fat grafting by Coleman in 1997. Since then, clinical practice using this natural filler is becoming more popular and the results are becoming more consistent. Nowadays autologous fat grafting is utilized broadly for both aesthetic and reconstructive purposes. With the beginning of the twenty first century, adipose stem cell (ASC) was discovered and regenerative medicine is facing a new era of evolution. ASC was applied to fat transfer and 'cell-assisted lipotransfer' technique could be developed. Eto et al. recently presented experimental results of fat graft survival. Their efforts disclosed the important role of ASCs in fat graft, and contributed the progress of fat transfer. Owing to the accumulation of knowledge related with fat graft survival, discovery of ASCs and advancements in surgical techniques, such as Coleman technique and cell-assisted lipotransfer, survival rate of the grafted fat increased and side effects such as fat necrosis decreased. Consequently, there is a new surgical trend of applying large volume fat grafting for augmentation mammoplasty and breast reconstruction with or without silicone implant. Recently, fat grafting is expanding it's limit to new field of treatment of burn, scar, and wound. This article reviews several significant advancements of fat grafting techniques in this century, furthermore intends to widen scientific understandings and contribute to be practiced as a feasible method.

The influence of platelet rich plasma gel applied to adipose-derived stem cells repair soft tissue wounds / 重庆医学

Objective To explore the role of ADSCs and PRP in soft tissue defect repairing .Methods Harvest adipose tissue from inguinalis fat pad of SD rats ,isolation ,culture ,and identification the ADSCs through three differentiation method .Take 30 male SD rats about 6-7 weeks old randomly .Randomly selected 12 rats been take blood from heart .Preparation the PRP with modi-fied appel method .To count platelet of whole blood and PRP under microscope .Take the remaining rats .Divided the rat into 3 groups(n=6 in each group) randomly ,of which group A treatment with ADSCs and PRP ;Group B treatment with ADSCs ;Group C treatment whit PRP .Selected one side of skin defect wound for test randomly and the relative side skin defect is for control ,Handle all of the control wound to group D ;Observe the grow th of granulation tissue on the wound surface ;observe the inflammatory sur-rounding and the degree about epithelial .statistical analysis and record the wound size ,and calculation the shrinkage rate of wound in different periods .Record the time of completely healing time .Histologic observation of the wound healing tissue .Results Plate-let counting showed platelet of PRP is 5 .21 times than whole blood .The wound completely healed time :group A (18 .25 ± 1 .44 ) days ,group B(19 .13 ± 1 .28)days ,group C(19 .72 ± 0 .87)days ,group D(22 .31 ± 1 .65) days ,The time of each treatment group and control group was significantly obvious(P< 0 .05) .At 3 ,7 ,11 and 15 days after experimental treatment ,compared with the control group the experiment group of wound contraction rate was significantly obvious (P<0 .05) .Conclusion Application of AD-SCs with PRP can enhance the quality and shorten the wound healing time than used them alone .

Brief Review of Adipose Derived Cells

Recent works state that adipose tissue hosts cells which are able to display various differentiation potentials. Moreover, this adult tissue is abundant and easy to sample with no ethic limitation. In addition, the simple isolation procedures provide a clear advantage for tissue engineering. The adipose cells which are used for tissue engineering can be isolated from the stromal vascular fraction (SVF) obtained after adipose tissue digestion that may be used either freshly prepared or after culture. In this last case, cultured cells represent a particular cell subpopulation, which is restricted to the adherent cell fraction of SVF, and termed adipose derived stromal cells(ADSCs). However, there is a confusing inconsistency in the literature in the use of terms to describe multipotent precursor cells from adipose tissue stroma, such as processed lipoasporate cells, ADSCs, preadipocytes, adipose stoma vascular cell fraction, SVF cells, and others. In addition, characteristics of such cells have not clearly been defined and still controversial. The aim of this brief and comprehensive review is to define terminologies for such cells, to describe preparation and isolation procedures for SVF cells and ADSCs, to summarize molecular characterization of SVF cells and ADSCs, and to discuss clinical cases using these cells.