Priming of adipose-derived stem cells with curcumin prior to cryopreservation preserves their functional potency: Towards an 'Off-the-shelf' therapy for burns.

Stem cells-based treatment for burn wounds require frozen cells as an off-the-shelf therapy; however, cryopreservation-induced oxidative stress resulted in post-thaw cell death or loss of cell functions, thus arrested their clinical practicality. Although antioxidant priming to stem cells increase their resistant to oxidative stress, but this strategy is still unexplored on cryopreserved cells. Herein, we investigated whether curcumin priming before cryopreservation could preserve the therapeutic potency of thawed stem cells. For this, unprimed and curcumin-primed adipose-derived stem cells (ASCs) were cryopreserved for one month. Post-thawing, cells were assessed for viability by trypan blue assay; metabolic activity by MTT assay; senescence by senescence-associated (SA)-ß-galactosidase activity staining assay; migration by scratch healing assay and; mRNA expression by real-time PCR. Subsequently, the healing potential was examined by injecting cells around the wound periphery of acidic burn in rats. Post-healing, skin architecture was histologically examined. Results demonstrated that, curcumin-primed frozen cells (Cryo/Cur-ASCs) showed better post-thaw viability, metabolic activity, migration ability and lower percent of senescence comparative to unprimed frozen cells (Cryo/ASCs). Curcumin priming alleviated the oxidative damage by activating the ROS-reducing cellular antioxidant system as shown by the evident increase in GSH levels and upregulated mRNA expression of glutathione peroxidase (GPx), superoxide dismutases (SOD1, SOD2), and catalase (CAT). Further, invivo findings revealed that Cryo/Cur-ASCs-treated wounds exhibited earlier wound closure with an improved architecture comparative to Cryo/ASCs and depicted healing capacity almost similar to Fresh/ASCs. Our findings suggested that curcumin priming could be effective to alleviate the cryo-induced oxidative stress in post-thawed cells.

Sebaceous carcinoma of forehead: A rare malignant lesion.

Sebaceous carcinoma is an uncommon malignant tumour of the sebaceous glands. This lesion typically occurs as painless and slow growing nodule in the eyelid region. In rarity, it can develop on buccal mucosa, head and neck, and other regions of the body in people in their 60s and 70s. Sebaceous carcinoma is locally aggressive and carries the potential of regional and distant spread. We present the case of a 15-year-old male who was diagnosed with sebaceous carcinoma over the forehead. After discussing the case in the board meeting, surgery was performed to remove the tumour with 1 cm margin. Outer table of the frontal bone was also removed and intraoperative frozen section was done for margin clearance. After the excision, soft tissue coverage was done with free Anterolateral thigh flap and the patient received six cycles of post-operative radiation therapy.

Cassia Angustifolia Primed ASCs Accelerate Burn Wound Healing by Modulation of Inflammatory Response.

BACKGROUND: Thermal traumas impose a huge burden on healthcare systems. This merits the need for advanced but cost-effective remedies with clinical prospects. In this context, we prepared a regenerative 3D-construct comprising of Cassia angustifolia extract (SM) primed adipose-derived stem cells (ASCs) laden amniotic membrane for faster burn wound repair. METHODS: ASCs were preconditioned with SM (30 µg/ml for 24 h), and subsequently exposed to in-vitro thermal injury (51 °C,10 min). In-vivo thermal injury was induced by placing pre-heated copper-disc (2 cm diameter) on dorsum of the Wistar rats. ASCs (2.0 × 105) pre-treated with SM (SM-ASCs), cultured on stromal side of amniotic membrane (AM) were transplanted in rat heat-injury model. Non-transplanted heat-injured rats and non-heat-injured rats were kept as controls. RESULTS: The significantly upregulated expression of IGF1, SDF1A, TGFß1, VEGF, GSS, GSR, IL4, BCL2 genes and downregulation of BAX, IL6, TNFα, and NFkB1 in SM-ASCs in in-vitro and in-vivo settings confirmed its potential in promoting cell-proliferation, migration, angiogenesis, antioxidant, cell-survival, anti-inflammatory, and wound healing activity. Moreover, SM-ASCs induced early wound closure, better architecture, normal epidermal thickness, orderly-arranged collagen fibers, and well-developed skin appendages in healed rat-skin transplanted with AM+SM-ASCs, additionally confirmed by increased expression of structural genes (Krt1, Krt8, Krt19, Desmin, Vimentin, α-Sma) in comparison to untreated-ASCs laden-AM transplanted in heat injured rats. CONCLUSION: SM priming effectively enabled ASCs to counter thermal injury by significantly enhancing cell survival and reducing inflammation upon transplantation. This study provides bases for development of effective combinational therapies (natural scaffold, medicine, and stem cells) with clinical prospects for treating burn wounds.

Protective and proliferative effect of Aesculus indica extract on stressed human adipose stem cells via downregulation of NF-κB pathway.

Inflammatory microenvironment after transplantation affects the proliferation and causes senescence of adipose-derived mesenchymal stem cells (hADMSCs) thus compromising their clinical efficacy. Priming stem cells with herbal extracts is considered very promising to improve their viability in the inflammatory milieu. Aesculus indica (A. indica) is used to treat many inflammatory diseases in Asia for decades. Herein, we explored the protective role of A. indica extract on human adipose-derived Mesenchymal Stem Cells (hADMSCs) against Monosodium Iodoacetate (MIA) induced stress in vitro. A. indica ameliorated the injury as depicted by significantly enhanced proliferation, viability, improved cell migration and superoxide dismutase activity. Furthermore, reduced lactate dehydrogenase activity, reactive oxygen species release, senescent and apoptotic cells were detected in A. indica primed hADMSCs. Downregulation of NF-κB pathway and associated inflammatory genes, NF-κB p65/RelA and p50/NF-κB 1, Interleukin 6 (IL-6), Interleukin 1 (IL-1ß), Tumor necrosis factor alpha (TNF-α) and matrix metalloproteinase 13 (MMP-13) were observed in A. indica primed hADMSCs as compared to stressed hADMSCs. Complementary to gene expression, A. indica priming reduced the release of transcription factor p65, inhibitory-κB kinase (IKK) α and ß, IL-1ß and TNF-α proteins expression. Our data elucidates that A. indica extract preconditioning rescued hADMSCs against oxidative stress and improved their therapeutic potential by relieving inflammation through regulation of NF-κB pathway.

Curcumin preconditioned human adipose derived stem cells co-transplanted with platelet rich plasma improve wound healing in diabetic rats.

AIM: Inflammatory and oxidative microenvironment at diabetic' wound site hinder the therapeutic efficacy of cell-based therapies in diabetic patients. The purpose of this study is to explore the competence of curcumin preconditioned human adipose derived cells (hASCs) in combination with platelet rich plasma (PRP) for the repair of wounds in diabetic rats. MAIN METHODS: The cytoprotective effect of curcumin preconditioning for hASCs against hyperglycemic stress was evaluated through analysis of cell morphology, viability, cytotoxicity, senescence, and scratch wound healing assays. Subsequently, the healing capacity of curcumin preconditioned hASCs (Cur-hASCs) added to PRP was examined in excisional wounded diabetic rat model. Healed skin biopsies were excised to analyze gene and protein expression of wound healing markers by qPCR and western blotting. Histopathological changes were observed through hematoxylin and eosin staining. KEY FINDINGS: We found that Cur-hASCs counteract the glucose stress much better than non-preconditioned hASCs by maintaining their cellular morphology and viability as well as metabolic potential. Further in vivo results revealed that, Cur-hASCs co-injected with PRP resulted in faster wound closure, improved fibroblast proliferation, increased neovascularization, marked reduction in inflammatory cells, and compact extracellular matrix with completely covered thick epithelium. Moreover, Cur-hASCs + PRP treatment significantly improved the expression of key healing markers such as pro-angiogenic (Vegf), dermal matrix deposition (Col1α1), cell migration (bFgf) and cell proliferation (Pcna) at wound site. SIGNIFICANCE: Our findings propose a combinatorial therapy (Cur-hASCs + PRP) as a novel modality to improve the efficacy of hASCs-based therapy for diabetic wounds.

Complex Reconstruction of Nose and Upper Lip after Assault.

A 21-year male, farmer by profession, was admitted in Plastic Surgery Department via outpatient department, with amputation of nose and upper lip with the knife during resistance against robbery attempt. He was having the blockage of the right side of nostril and difficulty in mouth opening. Upper lip vermilion and mucosal reconstruction with the bilateral facial artery musculo-mucosal (FAMM) flap and moustache reconstruction was done with visor flap. Split thickness skin graft (STSG) was done over the donor site. Division and insetting of visor flap was done after two weeks. After three months, all three nasal layers were reconstructed. The inner lining was reconstructed with the turn down flap, L-strut from rib cartilage, and the outer lining with forehead flap.

Oropharyngeal, Hypopharyngeal and Cervical Esophageal Reconstruction: An Experience of Pedicle Flaps.

OBJECTIVE: To review the experience of single-stage reconstruction following pharygolaryngectomy and cervical esophageal defect with pedicle flaps in a tertiary care centre. STUDY DESIGN: Retrospective study. PLACE AND DURATION OF STUDY: Jinnah Burn and Reconstructive Surgery Centre, Lahore, from June 2007 to June 2017. METHODOLOGY: All patients who underwent oropharyngeal, hypopharyngeal and cervical esophageal reconstruction with pedicled flaps were included. RESULTS: Thirty-two reconstructions were done, of which 16 (50.0%) were supraclavicular flaps, 12 (37.5%) were pectoralis major myocutaneous flaps (PMMF) and 4 (12.5%) platysma myocutaneous flaps (PMF). Among these, 24 (75%) were males and 8 (25%) females. The mean hospital stay was 18.75 +5.45 days. Complications were noted in 12 (37.5%), salivary fistula being the most frequent found in 6 (18.75%). Wound dehiscence was noted in 4 (12.5%), partial flap necrosis and wound infection present in 1 (3.1%) each. CONCLUSION: Oropharyngeal, hypopharyngeal and cervical esophageal reconstruction with pedicle flaps is still very useful and safe to perform, associated with minimal flap and donor site complications.

Efficacy Of Micro-Needling On Post Acne Scars.

BACKGROUND: Acne effects 80-90% of teenage population all around the world. Resulting scars can lead to facial disfigurement and psychological issues in affected population. To counter this problem many treatment options have been tried including resurfacing lasers, dermabrasion, peeling, fillers, platelet rich plasma therapy etc. All have shown variable results. Among these modalities, micro needling is showing promising results in treatment of acne scars due to collagen induction. This study was conducted to assess efficacy and safety of micro-needling on acne scars. METHODS: In this cross-sectional study a total of 50 patients (female 35, male 15) underwent treatment for post acne scarring, 4 sessions of micro-needling were done under local anaesthesia 3 weeks apart over the span of 2 months. Initial and follow up qualitative assessment was done. Photographs were taken of each patient on their subsequent visit, they were analysed and compared for final results. Grading was done by using Goodman Baron scale. RESULTS: 70% (35) were females and 30% (15) were males with mean age of 27.31±4.41 ranging from 19-35 years. After the treatment of scars, 73% (08) of Grade 4 scars have showed improvement by 2 grades, remaining 27% (03) showed improvement to Grade 3. In 20 patients with Grade 3 scars, 30% (06) showed improvement by 1 grade. Remaining 70% (14) improved by 2 grades. All patients with Grade 2 scar downgraded to grade 1 after treatment. Chi-square test was used to assess pre-treatment and post treatment grading improvement among subjects and was statistically significant. (X2=30.010 p=000).. CONCLUSIONS: Micro needling is an effective tool for aesthetic improvement of post acne scarring.

Comparison of efficacy and safety of intralesional triamcinolone and combination of triamcinolone with 5-fluorouracil in the treatment of keloids and hypertrophic scars: Randomised control trial.

The treatment of keloid and hypertrophic scar is challenging with no universally accepted mode for permanent ablation. Conventional therapies yield unpredictable results, significant complications and require elaborate hardware. OBJECTIVE: The objective was to establish the safety and efficacy of intralesional 5-fluorouracil (5-FU) for the treatment of keloids and hypertrophic scars. STUDY DESIGN: Randomized controlled trial (RCT). PLACE AND DURATION: It was conducted at the Jinnah Burn and Reconstructive Surgery Center/Allama Iqbal Medical College, Lahore, Pakistan from May 2012 to March 2013. SUBJECTS AND METHODS: We included 120 patients divided in two groups. The group A patients received intralesional triamcinolone acetonide (TAC) and the group B patients received both 5-FU and TAC. 8 injections at a week interval were given and patients were evaluated at the start of treatment and then at 4th and at 8th week during the treatment and then 4 weeks after the end of treatment. Patents were assessed for mean reduction in scar height, efficacy and complications. RESULTS: Total of 108 patients completed the study. The mean reduction in the scar height in group B (5-FU+TAC) 1.144+.4717 was markedly better than that of group A (TAC alone) 1.894+1.0751 (t=4.781, p=.000). The efficacy (defined previously as >50% reduction in initial scar height) was superior in group B 44 (77.2%) than that of group A 25 (49.0% (X2=9.260, p=.002). Recurrence was seen in 39.2% (20) of patients of the group A while in only 17.5% (10) of the cases of group B (P=0.012). Mean follow up was of 22 months. CONCLUSION: 5-FU+TAC is safe, easy to administer and effective treatment for problematic scars and has the lower rate of recurrence on larger follow up.

Extended Islanded Reverse Sural Artery flap for Staged Reconstruction of Foot Defects Proximal to Toes.

Obective:To assess the outcome of extended delayed reverse sural artery flap for reconstruction of foot defects proximal to toes in terms of flap survival, complication and extended area. STUDY DESIGN: Case series. PLACE AND DURATION OF STUDY: Jinnah Burn and Reconstructive Surgery Centre, Lahore, from February 2015 to April 2017. METHODOLOGY: Cases who underwent delayed sural artery flap were inducted. Preoperative hand-held doppler was done to confirm the location of perforator. Two suitable perforators were chosen to raise the extended flap by crossing the proximal limit in all cases. The pedicle was kept minimum 3 cm wide and perfusion was assessed. Flap was delayed for one week and vaccum-assisted closure (VAC) dressing was applied over wound. The second surgery was performed after one week. Proximal perforator was clamped and ligated after checking adequate perfusion of flap. Flap was insetted into defect. RESULTS: Thirty-two patients were reconstructed with delayed reverse sural artery flap. The mean age of the patients was 26.5 12.2 years. Twenty-four (75%) patients were males and 8 (25%) were females. Twenty-two (68.7%) cases were degloving wounds after road traffic accidents (RTA), 6 (18.7%) were diabetic foot wounds, 4 (12.5%) sustained injury after falling from height and 7 (21.8%) patients had fracture of metatarsals. Twenty-eight flaps were transferred after one week delay, and only in 4 cases, flap were transferred after two weeks. All flaps survived completely. Complications of infection noted in 3 (9.3%) flaps, 3 (9.3%) flaps showed tip necrosis, 2 (6.2%) flaps undergone epidermolysis and only 2 (6.2%) showed venous congestion. CONCLUSION: Delayed islanded reverse sural artery perforator flap is a reliable and versatile option for resurfacing soft tissue defects of lower limb proximal to the toes with lesser complications and extended coverage area.

Quantification and Further Refinements of Dorsal Metacarpal Artery Perforator Flap for Reconstruction of Wounds of Fingers upto Distal Interphalangeal Joints.

OBJECTIVE: To assess the outcome of dorsal metacarpal artery perforator flap for coverage of finger defects extending upto distal interphalangeal joint (DIPJ). STUDY DESIGN: Case series. PLACE AND DURATION OF STUDY: Jinnah Burn and Reconstructive Surgery Centre, Lahore, from March 2015 to May 2017. METHODOLOGY: Our study was carried out in two parts. 1st part of study was to measure average flap length in our population. Five hundred cases were enrolled to measure flap length, from pivot point of the flap to the distal border of extensor retinaculum. This length was traced to fingers to determine its coverage area. This was followed by clinical study in 35 cases. All patients with wounds over dorsal surface of fingers upto distal interphalangeal joint and volar surface of fingers upto mid of middle phalanx, single or multiple finger defects with exposed tendon joints or bones were included in the study. Patients with history of trauma to the dorsum of hand, metacarpal head or neck fracture and patients with history of diabetes or peripheral vascular disease were excluded. RESULTS: Flap length decreased from radial to ulnar side of hand. Average length of flap based on the second metacarpal artery was 7cm while of the third was 6.6 cm and the fourth was 6.1 cm. This flap length covered upto mid of middle phalanx in border digits while upto PIPJ in central digits. This data was confirmed in 35 patients in which 36 flaps were raised to cover finger defects. Thirty-four flaps survived completely while tip necrosis was seen in 2 cases. CONCLUSION: The dorsal metacarpal artery perforator flap is a thin, pliable flap, which has minimal donor-site morbidity. It can reliably cover soft tissue defects of dorsum of fingers upto mid of middle phalanx in border digits and upto PIPJ in central digits.