Efficacy of mesenchymal stem cell-delivery using perpendicular multi-needle injections to the skin: Evaluation of post-ejection cellular health and dermal delivery.

AIM: Mesenchymal stem cell (MSC)-therapy is increasingly being evaluated in clinical trials. Dermal delivery is not only time consuming but also unreliable, potentially hampering the therapeutic result. Therefore, qualification of cell delivery protocols is essential. This study evaluated a clinically relevant automated multi-needle injection method for cutaneous MSC-therapy, allowing the skin to be readily and timely treated, by assessing both the cellular health post-ejection and dermal delivery. METHODS: Following dispensation through the injector (31 G needles: 9- or 5-pin) the cellular health and potency (perceived- and long-term (12 h) viability, recovery, metabolism, adherence, proliferation and IDO1-expression) of adipose-derived stem cells (10-20-50 ×106 cells/ml) were assessed in vitro in addition to dermal delivery of solution in human skin. RESULTS: No significant detrimental effect on the perceived cell viability, recovery, metabolism, adherence or IDO1-expression of either cell concentration was observed. However, the overall long-term viability and proliferation decreased significantly regardless of cell concentration, nonetheless marginally. An injection depth above 1.0 mm resulted in all needles piercing the skin with dermal delivery from up to 89% needles and minimal reflux to the skin surface, and the results were confirmed by ultrasound and histology. CONCLUSION: The automated injector is capable of delivering dermal cell-doses with an acceptable cell quality.

Mesenchymal stem cell therapy of acute thermal burns: A systematic review of the effect on inflammation and wound healing.

AIM: Mesenchymal stem cell (MSC) therapies are emerging as a promising strategy to promote tissue repair, and may extend their utility to burn care. This comprehensive review of the extant literature, evaluated all in vivo studies, to elucidate the potential protective and therapeutic effect of MSCs in acute thermal skin burns. METHODS: PubMed was systematically searched, according to PRISMA guidelines, and all relevant preclinical and clinical studies were included according to pre-specified eligibility criteria. RESULTS: Forty-two studies were included in a qualitative synthesis, of which three were human and 39 were animal studies. The preclinical studies showed that MSCs can significantly reduce inflammation, burn wound progression and accelerate healing rate of acute burns. The underlying mechanisms are complex and not fully understood but paracrine modulators, such as immunomodulatory, antioxidative and trophic factors, seem to play important roles. Allogeneic MSC therapy has proved feasible in humans, and could allow for prompt treatment of acute burns in a clinical setting. CONCLUSION: MSC therapy show positive results, regarding improved burn wound healing and immunologic response. However, most findings are based on small animal studies. Randomized clinical trials are warranted to investigate the regenerative effects in human burns before translating the findings into clinical practice.

Cryopreservation of adipose-derived stromal/stem cells using 1-2% Me2SO (DMSO) in combination with pentaisomaltose: An effective and less toxic alternative to comparable freezing media.

Mesenchymal stromal/stem cells (MSCs) derived from bone marrow, umbilical cord and especially adipose tissue are increasingly being explored for their therapeutic potential to treat a wide variety of diseases. A prerequisite for most allogeneic off-the-shelf and some autologous MSC therapies is the ability to safely and efficiently cryopreserve cells during production or for storage prior to treatment. Dimethyl sulfoxide (Me2SO) is still the commonly used gold standard cryoprotectant (CPA). However, undesirable cellular impacts and side effects of Me2SO have led to an increasing demand for the development of safe and effective alternatives. This study investigated the effect of pentaisomaltose as a CPA for cryopreservation of adipose-derived stromal/stem cells (ASCs). We compared pentaisomaltose-based freezing media containing 1% Me2SO (PIM1) or 2% Me2SO (PIM2) to our in-house freezing media formulation containing 10% Me2SO (STD10) and to CryoStor freezing media containing 2% or 10% Me2SO (CS2 and CS10). We assessed the recovery of viable ASCs, their phenotype, differentiation potential, proliferation potential, and migratory potential. Further, their immunomodulatory potential was assessed by measuring their ability to suppress T cell proliferation and express immunomodulatory markers. The results showed that the post-thaw viability of ASCs cryopreserved with STD10, CS10 and PIM2 was improved compared to that of CS2. The recovery of ASCs with PIM1 and PIM2 was also improved compared to that of CS2. Proliferation and migration were comparable among the tested freezing media. The results showed no difference in the induction of PDL1, PDL2 or IDO1 expression. Nevertheless, the potential of cryopreserved ASCs to suppress T cell proliferation was reduced when the Me2SO concentration was reduced (CS10>STD10>CS2 and PIM2>PIM1). Altogether, the migratory and immunomodulatory potential combined with improved recovery indicate that the addition of pentaisomaltose in the freezing media may allow for the reduction of the Me2SO concentration to 2% while retaining a more potent cell product that what is recovered using comparable freezing media. With the desire to reduce the amount of Me2SO, these results suggest that 2% and potentially even 1% Me2SO in combination with 10% pentaisomaltose could be an effective and less toxic alternative to comparable freezing media.

Cell-Enriched Fat Grafting Improves Graft Retention in a Porcine Model: A Dose-Response Study of Adipose-Derived Stem Cells versus Stromal Vascular Fraction.

BACKGROUND: Cell-enrichment of fat grafts has produced encouraging results, but the optimal concentrations and types of added cells are unknown. The authors investigated the effects of enrichment with various concentrations of ex vivo-expanded adipose-derived stem/stromal cells and stromal vascular fraction on graft retention in a porcine model. METHODS: Adipose-derived stem/stromal cells were culture-expanded, and six fat grafts (30 ml) were prepared for each minipig (n = 13). The authors investigated grafts enriched with 2.5 × 10 to 20 × 10 adipose-derived stem cells/ml and stromal vascular fraction and nonenriched control grafts. Each pig served as its own control. Magnetic resonance imaging was performed immediately after grafting and 120 days postoperatively before the pigs were euthanized, and histologic samples were collected. RESULTS: The authors recorded an enhanced relative graft retention rate of 41 percent in a pool of all cell-enriched grafts compared to the nonenriched control (13.0 percent versus 9.2 percent; p = 0.0045). A comparison of all individual groups showed significantly higher graft retention in the 10 × 10-adipose-derived stem/stromal cells per milliliter group compared with the control group (p = 0.022). No significant differences were observed between the cell-enriched groups (p = 0.66). All fat grafts showed a significantly better resemblance to normal fat tissue in the periphery than in the center (p < 0.009), but no differences in overall graft morphology were observed between groups (p > 0.17). CONCLUSIONS: Cell-enriched fat grafting improved graft retention and was feasible in this porcine model. No significant differences in graft retention were observed among the various adipose-derived stem/stromal cell concentrations or between adipose-derived stem/stromal cell and stromal vascular fraction enrichment. Future studies using this model can help improve understanding of the role of adipose-derived stem/stromal cells in cell-enriched fat grafting.

[Necrobiotic xanthogranuloma misdiagnosed as a planocellular carcinoma].

In this case report, an 89-year-old woman presented with a subcutaneous tumour in the orbital region. Fine needle aspiration showed no signs of malignancy. Following progressive tumour growth and cutaneous manifestation, an incisional biopsy was taken, which showed squamous cell carcinoma. The tumour was excised, and the patho-logical report showed necrobiotic xanthogranuloma being a rare histiocytic disease characterised by slow growth of nodules and plaques. Because it is rare, it can pose a diagnostic challenge, which is why, this report is an attempt to shed light on the disease and to aid its diagnosis.

[Breast abnormalities in women].

Breast abnormalities in women are common and may have physical as well as psychological consequences. Many of the conditions can be treated surgically. It is important, that the patients are correctly diagnosed and referred to the appropriate department for further evaluation and treatment. This review discusses the most common breast abnormalities to ensure awareness among general practitioners and correct referral for treatment.

[Treating scars with lipofilling].

Hypertrophic, keloidal and contracted scars can be cosmetically and functionally debilitating, and by patients this is often perceived as traumatising. Psychological and social impairments are common in these types of scars. Currently, autologous lipofilling, with or without enrichment with stem cells, is performed to improve scar quality. In this review, we evaluate the evidence on lipofilling as a treatment of scars. The literature mainly consists of studies of low-level evidence, i.e. they lack controls and randomisation. Nevertheless, all studies indicate, that lipofilling may improve the quality of scars.

Surgical treatment of disabling conditions caused by anogenital lichen sclerosus in women: An account of surgical procedures and results, including patient satisfaction, benefits, and improvements in health-related quality of life.

BACKGROUND: Anogenital lichen sclerosus (LS), a chronic dermatitis that causes scarring and introital stenosis, may prevent sexual intercourse and reduce health-related quality of life (QoL). Surgery can restore the anatomy, allowing patients to resume their sexual lives. This study investigates outcomes in women treated with local skin flaps. METHODS: Thirty-eight consecutive LS-verified patients, surgically treated for debilitating conditions between 1990 and 2013, were retrospectively evaluated. A survey measured patient satisfaction, benefits, and health-related QoL, and the patients were also evaluated by a long-term clinical follow-up. RESULTS: In total, 33 patients (87%) experienced dyspareunia, 24 of whom could not perform coitus. At mean short-term follow-up (10.0 months), only five patients (15%) reported dyspareunia; for seven patients, the outcome was unknown. The survey response rate was 87%, and the mean time from treatment to response was 7.6 years. Twenty of 24 patients reported dyspareunia. Seventy-five percent of patients with preoperative dyspareunia reported a surgical benefit, 74% were satisfied/very satisfied with the cosmetic and overall results, respectively, and 58% reported that surgery had improved their sexual lives. The mean long-term clinical follow-up was 8.4 years. The follow-up rate was 78%. The main reason for recurrent dyspareunia was minor LS relapse (50%); these patients were still able to have coitus, and dyspareunia was reported as considerably minor compared to before surgery; 38% had more severe LS relapse, resulting in apareunia. CONCLUSIONS: Surgery for LS sequelae provides acceptable short-term functional results, enabling patients to resume coitus, with high patient satisfaction reported. However, the chronic relapsing nature of LS consequently provides varying and often short-term coital improvements following surgery.