Culturing Adipose-Derived Stem Cells Under Serum-Free Conditions.

Growing adipose-derived stem cells (ADSC) in serum-free conditions is important as it represents a way of expanding multipotent cells in a clinical grade medium. Most cultured ADSC are expanded and tested in serum-containing media, which can pose significant health risks if these cells were used in clinical applications. Moreover, cells grown in serum-free conditions behave very different than those cultured in serum-containing media. Here, we present a technique to culture adipose-derived stem cells in serum-free conditions. The methods described in this chapter were optimized for ovine ADSC. The appropriate optimization should be done for other cell lines.

Oral bacteriome of HIV-1-infected children from Rio de Janeiro, Brazil: Next-generation DNA sequencing analysis.

BACKGROUND AND AIM: This study compared the oral bacteriome between HIV-1-infected and non-HIV-1-infected Brazilian children/teenagers. METHODS: Whole saliva, biofilm from the dorsal surface of the tongue and biofilm from supragingival and subgingival sites were collected from 27 HIV-1-infected and 30 non-HIV-1-infected individuals. Bacterial genomic DNA was extracted and 16S rRNA genes were sequenced using next-generation sequencing technology (Ion Torrent). RESULTS: In the supragingival biofilm, the phylum Firmicutes and genus Streptococcus sp. were more frequent in HIV-1-infected (95% and 78%, respectively) than in non-HIV-1-infected individuals (40% and 24%, respectively). In the subgingival biofilm of HIV-infected participants, the relative abundance of the Veillonella sp. and Prevotella sp. genera were higher than in non-HIV-1-infected participants. On the tongue, the genera with greater relative abundance in HIV-1-infected individuals were Neisseria sp. (21%). In saliva, the difference of the genus Prevotella sp. between non-HIV-1-infected and HIV-1-infected individuals was 15% and 7%, respectively. The Chao index revealed an increase in the richness of both sub- and supragingival biofilms in the HIV-1-infected samples compared with non-HIV-1-infected samples. CONCLUSION: HIV-1-infected children/teenagers have a higher frequency of the phyla Firmicutes and genus Streptococcus, and their oral microbiome shows more complexity than that of non-HIV-1-infected children/teenagers.

Effect of Titanium Surfaces on the Osteogenic Differentiation of Human Adipose-Derived Stem Cells.

PURPOSE: To investigate whether human adipose-derived stem cells will form a functional bone-like matrix on titanium substrates. The behavior of human adipose-derived stem cells was examined when grown in either serum-free, xeno-free stem cell growth medium or osteogenic differentiation medium and cultured on either machined titanium (MTi) or on roughened alumina-blasted titanium (ABTi) discs. MATERIALS AND METHODS: Cellular proliferation, extracellular mineralized matrix production, osteogenic-related protein production (RUNX2 and osteocalcin), and gene expression for pluripotency and self-renewal (TERT and OCT4) and osteogenic-related (MSX2, RUNX2, and BGLAP) genes were performed. RESULTS: Human adipose-derived stem cells in serum-free medium (hADSC) proliferated at a higher rate compared with osteogenically differentiated cells (hOS-ADSC); however, the osteogenically committed cells produced more mineralized matrix on the titanium surfaces compared with either tissue culture plastic or the undifferentiated cells. The immunofluorescence analysis showed that human adipose-derived stem cells cultured in serum-free medium and osteogenic differentiation medium produced RUNX2 on both the machined titanium surface and on the alumina-blasted titanium surface after 7 days in culture. Only osteogenically differentiated cells produced osteocalcin after 21 days. Relative gene expression showed stable expression of MSX2, RUNX2, and BGLAP over time on all surfaces. Only osteogenically differentiated cells displayed osteogenic characteristics over time. CONCLUSION: This study confirmed that human adipose-derived stem cells could be successfully grown in serum-free, xeno-free culture medium suitable for clinical use. Adipose-derived stem cells thus show potential utility for bone regeneration in association with titanium surfaces.

Growing Adipose-Derived Stem Cells Under Serum-Free Conditions.

Growing adipose-derived stem cells (ADSC) in serum-free conditions is important as it represents a way of expanding multipotent cells in a clinical grade medium. Most cultured ADSC are expanded and tested in serum-containing media, which can pose significant health risks if these cells were used in clinical applications. Moreover, cells grown in serum-free conditions behave significantly different than those cultured in serum-containing media. Here, we present a technique to culture adipose-derived stem cells in serum-free conditions. The methods described in this chapter were optimized for ovine ADSC. The appropriate optimization should be done for other cell lines.

Nonprocessed adipose tissue graft in the treatment of peri-implant osseous defects in the rabbit's tibiae: a pilot study.

We hypothesized that a new technique using nonprocessed adipose tissue could regenerate bone around dental implants. Eighteen rabbits received 1 implant per tibia surrounded by a surgically created osseous defect. The defects were assigned for treatment into 3 groups: C, AT, and AB. The percentages of bone-to-implant contact were 17.64% ± 16.22% (AB), 3.54% ± 7.08% (AT), and 12.71% ± 10.11% (C) (ρ = 0.25). The use of adipose tissue around surgically created peri-implant osseous defects interferes with bone formation.

Nonprocessed adipose tissue graft in the treatment of dehiscence bone defects in rabbit tibiae: a pilot study.

OBJECTIVE: To evaluate the bone repair of surgically created dehiscence-type defects (3 × 5 mm) around dental implants in rabbit tibia using nonprocessed adipose tissue graft or autogenous bone graft. MATERIALS AND METHODS: The bone defects were randomly assigned to 3 groups: blood clot (C), autogenous bone (AB), and nonprocessed adipose tissue (AT). After 3 months, the animals were euthanized. Histomorphometric analyses were performed, and the results were analyzed with analysis of variance and Tukey tests (P ≤ 0.05). Statistics were performed for the percentage of the bone-to-implant contact (BIC) and bone area (BA) within the limits of the threads. RESULTS: The results for BIC in the AT (37.75% ± 28.03%) and C (40.57 ± 13.71%) groups were statistically similar, whereas the AB group had the greatest percentage of BIC (83.37% ± 11.85%). For all groups, the BA percentage was similar (61.48% ± 30.89% in AT; 72.90% ± 14.10% in C; 84.23% ± 11.96% in AB), with no statistically significant differences. CONCLUSION: Nonprocessed adipose tissue is not a comparable substitute for autogenous bone in the treatment of dehiscence bone defects around titanium dental implants.