Effect of cold plasma on periodontal wound healing-an in vitro study.

OBJECTIVES: Cold atmospheric plasma (CAP), a room temperate ionized gas, seems to be a possible way to enhance tissue recovery. An in vitro study was conducted to investigate the influence of medical CAP on the regenerative capacity of human periodontal ligament (PDL) cells. MATERIAL AND METHODS: Human PDL cells were subjected to CAP at various intensities, distances, and durations. The effects of CAP on a number of specific markers were studied at transcriptional level using real-time PCR. Additionally, an in vitro wound healing assay was applied to PDL cell monolayers either in the presence or absence of CAP by using JuLI™ Br Live Cell Analyzer and software. Finally, cell viability of CAP-treated cells was analyzed by an XTT assay. RESULTS: CAP treatment enhanced significantly the expression of the cytokines tumor necrosis factor (TNF)α, cyclooxygenase (COX)2, interleukin (IL)-1ß, IL-6, IL-8, collagen (COL)1α, and matrix metalloproteinase (MMP)1, as well as the proliferation markers Ki67 and proliferating cell nuclear antigen (PCNA), but downregulated apoptotic markers Apaf1 and p53. Additionally, the in vitro wound healing rate was significantly enhanced after CAP application. Moreover, CAP treatment resulted in a significantly increased cell viability in the XTT assay. CONCLUSION: This in vitro study shows that CAP has regulatable effects on markers of periodontal wound healing thereby underlining the potential use of CAP as a benefit treatment strategy. CLINICAL RELEVANCE: Our study demonstrates the application of CAP in the treatment of oral pathologies suggesting a promising future treatment approach.

The micromass formation potential of human adipose-derived stromal cells isolated from different various origins.

BACKGROUND: Adult stem cells appear to be a promising subject for tissue engineering, representing an individual material for regeneration of aged and damaged cells. Especially adipose derived stromal cells (ADSC), which are easily to achieve, allow an encouraging perspective due to their capability of differentiating into miscellaneous cell types. Here we describe the in vitro formation of human subcutaneous, visceral and omental ADSC micromasses and compare their histological attributes while being cultivated on collagen membranes. METHODS: Subcutaneous, visceral and omental fat tissue derived cells were isolated and processed according to standard protocols. Positively stained cells for CD13, CD44 and CD90 were cultivated on agarose in order to study micromass formation using a special method of cell tracking. Stained paraffin-embedded micromasses were analysed morphologically before and after being plated on collagen membranes. RESULTS: The micromass formation process was similar in all three tissue types. Subcutaneous fat tissue derived micromasses turned out to develop a more homogeneous and compact shape than visceral and omental tissue. Nevertheless all micromasses adhered to collagen membranes with visible spreading of cells. The immune histochemical (IHC) staining of subcutaneous, visceral and omental ADSC micromasses shows a constant expression of CD13 and a decrease of CD44 and CD 90 expression within 28 days. After that period, omental fat cells don't show any expression of CD44. CONCLUSION: In conclusion micromass formation and cultivation of all analysed fat tissues can be achieved, subcutaneous cells appearing to be the best material for regenerative concepts.

Regenerative potential of human adipose-derived stromal cells of various origins.

In regenerative concepts, the potential of adult stem cells holds great promise concerning an individualized therapeutic approach. These cells provide renewable progenitor cells to replace aged tissue, and play a significant role in tissue repair and regeneration. In this investigation, the characteristics of different types of adipose tissue are analysed systematically with special attention to their proliferation and differentiation potential concerning the angiogenic and osteogenic lineage. Tissue samples from subcutaneous, visceral, and omental fat were processed according to standard procedures. The cells were characterized and cultivated under suitable conditions for osteogenic and angiogenic cell culture. The development of the different cell cultures as well as their differentiation were analysed morphologically and immunohistochemically from cell passages P1 to P12. Harvesting and isolation of multipotent cells from all three tissue types could be performed reproducibly. The cultivation of these cells under osteogenic conditions led to a morphological and immunohistochemical differentiation; mineralization could be detected. The most stable results were observed for the cells of subcutaneous origin. An osteogenic differentiation from adipose-derived cells from all analysed fatty tissues can be achieved easily and reproducibly. In therapeutic concepts including angiogenic regeneration, adipose-derived cells from subcutaneous tissue provide the optimal cellular base.