Application to Quantify Ulcer Areas and Track Their Progress.

Chronic ulcer treatments, such as those for diabetic foot ulcers and pressure sores, require prolonged treatment periods. Availability of effective objective indicators to determine treatment method efficacy is limited. Ulcer area is the agreed-upon indicator for ulcer healing because it contracts and/or undergoes epithelialization as healing occurs. Ulcer surface properties such as healthy or infected granulation and slough or necrotic tissue are also used. This study aimed to develop a user-friendly application automating the ulcer area measurement process and included a graphical time-series display of ulcer components manually classified by users. Images of ulcers photographed with adjacent circular 1.5-cm diameter stickers were prepared. In the application, users manually categorized and color-coded each image into five component types based on different ulcer characteristics. The application calculated the area of each component in pixels and then estimated the actual area using the sticker area as a reference. It also collated color-coded images and presented graphical illustrations of changes in area over time. The results indicated the application successfully automated area measurements of each ulcer component and graphical displays of changes in ulcer component areas over time. It enabled users to visually track quality changes and the chronic ulcer healing process. Historically, ulcer assessments are subjectively conducted via visual examination by physicians, creating less reproducible, objective data. Although ulcer properties still required manual entry by users, our application streamlined ulcer area measurement and time-course visualization and sets the groundwork for a fully automated artificial-intelligence-driven ulcer diagnosis system.

Pinwheel-Shaped Titanium Plates Should Be Fixed to the Skull Using All Screw Holes to Protect the Plates from Being Bent.

INTRODUCTION: In cranioplasty, pinwheel-shaped titanium mini plates are frequently used to cover bone defects produced by burr holes, and it is common to insert screws through only a few of the holes in cranial flap fixation. PRESENTATION OF CASE: A 69-year-old man who had undergone clipping surgery for subarachnoid hemorrhage 16 years previously visited our clinic because a titanium plate had penetrated his scalp one month after he was hit on the head by a wall cabinet. Imaging studies revealed that part of the titanium plate had bent outwards and penetrated the skin. The plate was surgically removed, a relief skin incision was made 6 cm posterior to the skin defect to suture the defected portion without causing tension, and a skin graft was applied to the relief skin incision portion. Two months after the maneuver, the skin graft had been successfully incorporated without infection. DISCUSSION: Even after the subcutaneous and the cutaneous tissue have completely covered the pinwheel-shaped titanium mini plate, an edge without screw fixation can be easily bent by a hard blow to the overlying scalp. We recommend fixation of pinwheel-shaped titanium plates used in cranioplasty through all screw holes to protect against the plate being bent.

Flap Blood Glucose as a Sensitive and Specific Indicator for Flap Venous Congestion: A Rodent Model Study.

BACKGROUND: Flap blood glucose decreases when flap congestion occurs. The hypothesis that flap blood glucose works as an indicator for venous congestion was tested experimentally, and flap congestion was reproduced in rodent models. METHODS: Blood glucose levels of a rat abdominal skin flap, with or without its vein pedicle clamped, were checked before and every 10 minutes after flap elevation. In rats whose pedicle vein was shut off, it was further followed up every 5 minutes after declamping. To examine the effect of systemic blood glucose on flap blood glucose, in some rats, glucose solution was administered intraperitoneally before the experiment to artificially produce hyperglycemia. Forty-two rats were divided into four groups, with (n = 24) or without (n = 18) venous blockage and with (n = 20) or without (n = 22) glucose preloading. RESULTS: Flap blood glucose decreased rapidly to off-scale low (<20 mg/dl) within 40 minutes only when the vein pedicle was shut off in normoglycemic (40 ± 8.2 minutes, mean ± SD) and hyperglycemic (40 ± 9.9 minutes) rat groups (p < 0.01). There was no significant difference in the time taken for the flap blood glucose to decrease to off-scale low after venous blockage between both groups (p = 0.379). When the vein was declamped, flap blood glucose again rapidly returned to the systemic level in 15 minutes or earlier in both groups (p = 0.0283). CONCLUSIONS: Flap blood glucose sensitively and specifically reflects the state of vein occlusion, whether the systemic blood glucose is normal or high. The authors' results indicate that flap blood glucose works as a reliable indicator for the venous system.

Evaluation of the In Vivo Kinetics and Biostimulatory Effects of Subcutaneously Injected Hyaluronic Acid Filler.

BACKGROUND: Because subcutaneously injected hyaluronic acid filler is absorbed over 6 months to 1 year after the treatment of facial wrinkles, frequent retreatment may be required. However, persistent long-term effects are often clinically observed when hyaluronic acid filler is injected as a bolus for facial augmentation. Therefore, the authors investigated, over time, the changes in volume and histologic features of subcutaneous bolus injections of hyaluronic acid. METHODS: Hyaluronic acid filler was subcutaneously injected as a bolus into the dorsum of 6-week-old rats. At several time points (immediately after injection and 4, 8, 16, 32, and 64 weeks thereafter), magnetic resonance imaging was introduced to observe morphologic changes and to measure volume. Histologic examination of sectioned tissues was also performed. RESULTS: The average volume increased for up to 4 weeks after injection and then gradually decreased, with 74.8 percent of the injected volume remaining after 64 weeks, with no statistical difference compared to the initial volume. Histologic analysis revealed that lattice structures were created by fibroblasts and collagen fibers, and blood vessels and adipocytes were also generated in the filler. CONCLUSIONS: Although subcutaneous bolus injections of hyaluronic acid filler exhibited flattening, the total volume was maintained even after 64 weeks. Histologically, hyaluronic acid filler acted as a scaffold for autogenous tissue replacement by means of fibroblast migration and proliferation, collagen induction, and angiogenesis, followed by proliferation of adipocytes. This study demonstrates that the total volume is maintained long-term by replacing part of the injected hyaluronic acid filler with autologous tissues.

The in vivo effect of esculetin ointment and esculetin-mixed Zyderm for Zyderm.

BACKGROUND: Injectable collagen is often used for treatment of wrinkles or scars in cosmetic surgery. However, it is degraded within a short period after subcutaneous injection. The authors aimed to achieve a long-lasting effect of the filler with a new collagenase inhibitor, esculetin (6,7-dihydroxy-2H-chromen-2-one). METHOD: Nude mice were divided into two study groups and a control group (35 mg cattle collagen): (1) those implanted with Zyderm 0.3 g subcutaneously into the dorsal region followed by daily topical application of 5% esculetin ointment (0.5 g/day) to the skin of the implanted area (the 5% esculetin ointment group), and (2) those implanted with a mixture of Zyderm 0.3 g and esculetin (1 to 4 mM) (the esculetin-mixed Zyderm groups). In each group, Zyderm was removed at different time points to measure the wet weight and hydroxyproline level. Furthermore, each removed Zyderm specimen was sectioned for histologic examination with Azan staining and immunostaining. RESULTS: In the esculetin ointment group and the 2 mM esculetin-mixed Zyderm group, the hydroxyproline levels at 30, 60, and 90 days were significantly higher than those in the control group, suggesting that esculetin suppresses the biodegradation of Zyderm. There was no significant difference in hydroxyproline level between the esculetin ointment group and the 2 mM esculetin-mixed Zyderm group; biodegradation occurred to a similar extent with either method of application. CONCLUSIONS: An atelocollagen implant is used as a safe and effective scaffold material for tissue regeneration. Future applications of the present study are expected.

Sphingosine-1-phosphate receptor-2 deficiency leads to inhibition of macrophage proinflammatory activities and atherosclerosis in apoE-deficient mice.

Sphingosine-1-phosphate (S1P) is a biologically active sphingolipid that has pleiotropic effects in a variety of cell types including ECs, SMCs, and macrophages, all of which are central to the development of atherosclerosis. It may therefore exert stimulatory and inhibitory effects on atherosclerosis. Here, we investigated the role of the S1P receptor S1PR2 in atherosclerosis by analyzing S1pr2-/- mice with an Apoe-/- background. S1PR2 was expressed in macrophages, ECs, and SMCs in atherosclerotic aortas. In S1pr2-/-Apoe-/- mice fed a high-cholesterol diet for 4 months, the area of the atherosclerotic plaque was markedly decreased, with reduced macrophage density, increased SMC density, increased eNOS phosphorylation, and downregulation of proinflammatory cytokines compared with S1pr2+/+Apoe-/- mice. Bone marrow chimera experiments indicated a major role for macrophage S1PR2 in atherogenesis. S1pr2-/-Apoe-/- macrophages showed diminished Rho/Rho kinase/NF-κB (ROCK/NF-κB) activity. Consequently, they also displayed reduced cytokine expression, reduced oxidized LDL uptake, and stimulated cholesterol efflux associated with decreased scavenger receptor expression and increased cholesterol efflux transporter expression. S1pr2-/-Apoe-/- ECs also showed reduced ROCK and NF-κB activities, with decreased MCP-1 expression and elevated eNOS phosphorylation. Pharmacologic S1PR2 blockade in S1pr2+/+Apoe-/- mice diminished the atherosclerotic plaque area in aortas and modified LDL accumulation in macrophages. We conclude therefore that S1PR2 plays a critical role in atherogenesis and may serve as a novel therapeutic target for atherosclerosis.

S1P(2), the G protein-coupled receptor for sphingosine-1-phosphate, negatively regulates tumor angiogenesis and tumor growth in vivo in mice.

Sphingosine-1-phosphate (S1P) has been implicated in tumor angiogenesis by acting through the G(i)-coupled chemotactic receptor S1P(1). Here, we report that the distinct receptor S1P(2) is responsible for mediating the G(12/13)/Rho-dependent inhibitory effects of S1P on Akt, Rac, and cell migration, thereby negatively regulating tumor angiogenesis and tumor growth. By using S1P(2)(LacZ/+) mice, we found that S1P(2) was expressed in both tumor and normal blood vessels in many organs, in both endothelial cells (EC) and vascular smooth muscle cells, as well as in tumor-associated, CD11b-positive bone marrow-derived cells (BMDC). Lewis lung carcinoma or B16 melanoma cells implanted in S1P(2)-deficient (S1P(2)(-/-)) mice displayed accelerated tumor growth and angiogenesis with enhanced association of vascular smooth muscle cells and pericytes. S1P(2)(-/-) ECs exhibited enhanced Rac activity, Akt phosphorylation, cell migration, proliferation, and tube formation in vitro. Coinjection of S1P(2)(-/-) ECs and tumor cells into wild-type mice also produced a relative enhancement of tumor growth and angiogenesis in vivo. S1P(2)(-/-) mice were also more efficient at recruiting CD11b-positive BMDCs into tumors compared with wild-type siblings. Bone marrow chimera experiments revealed that S1P(2) acted in BMDCs to promote tumor growth and angiogenesis. Our results indicate that, in contrast to endothelial S1P(1), which stimulates tumor angiogenesis, S1P(2) on ECs and BMDCs mediates a potent inhibition of tumor angiogenesis, suggesting a novel therapeutic tactic for anticancer treatment.

Digital artery perforator (DAP) flaps: modifications for fingertip and finger stump reconstruction.

UNLABELLED: Various fingertip reconstructions have been reported for situations where microsurgical finger replantation is impossible. One method is the digital artery perforator (DAP) flap. Herein we report 13 DAP flaps for fingertip and finger stump reconstruction following traumatic finger amputations, highlighting modifications to the originally described DAP flap. METHODS: From October 1998 to December 2007, a total of 13 fingers (11 patients) underwent fingertip and finger stump reconstruction with modified DAP flaps following traumatic finger amputations. We performed six adipocutaneous flaps, three adipose-only flaps, two supercharged flaps and two extended flaps. Flap size ranged from 1.44 to 8 cm(2) (average 3.25 cm(2)). RESULTS: All flaps survived completely with the exception of partial skin necrosis in two cases. One of these cases required debridement and skin grafting. Our initial three cases used donor-site skin grafting. The donor site was closed primarily in the 10 subsequent cases. No patients showed postoperative hypersensitivity of repaired fingertips. Semmes-Weinstein (SW) test result for flaps including a digital nerve branch did not differ from those without (average 4.07 vs. 3.92). CONCLUSIONS: Modified DAP flaps allow for preservation of digital length, volume and finger function. They can be raised as adiposal-only flaps or extended flaps and supercharged through perforator-to-perforator anastomoses. The donor defect on the lateral pulp can be closed primarily or by skin grafting. For traumatic fingertip and finger stump reconstructions, DAP flaps deliver consistent aesthetic and functional results.

New thoracodorsal artery perforator (TAPcp) flap with capillary perforators for reconstruction of upper limb.

BACKGROUND: Thoracodorsal artery perforator (TAP) flap is not yet popularised because the dominant large muscle perforators are often absent. Even in those cases, small capillary perforators exist around the proximal portion of the lateral border of the latissimus dorsi muscle, and they have a potential of a large skin territory. To overcome the weakness of thoracodorsal artery muscle perforator (TAP-MCp) and septocutaneous perforator (TAP-SCp) flaps, we present a new TAP flap with capillary perforators (TAPcp) flap. METHODS AND RESULTS: A total of 14 patients with upper-limb defects were repaired with free TAPcp flap. Among them were three combined TAP flaps with vascularised scapula bone flap and eight flow-through flaps. Recipient sites were one axilla, three upper arms, one elbow, two forearms and seven hands. The only postoperative complication was a partial necrosis of the flaps. CONCLUSION: A new TAP flap with capillary perforators is very useful for the reconstruction of upper limb. The advantage is easier elevation within short time (does not require intramuscular dissection) with long or short vascular pedicle. The flap can be elevated in a supine position, and even lateral descending branch can be a pedicle vessel. The flap can be a flow-through flap and is less invasive, because the remaining muscle can be preserved along with the motor nerve.

Intravascular stenting (IVaS) method for fingertip replantation.

Remarkable progress has been made in microsurgery. However, fingertip replantation following amputation has not gained much popularity because of its technical difficulty. We have developed the intravascular stenting (IVaS) method, in which a nylon monofilament is placed inside the vessel lumen to act as a temporary stent, facilitating anastomosis completion. This report describes 7 fingertip replantations using the IVaS method. Intravascular stent size varied from 4-0 to 6-0 (0.199-0.07 mm diameter). There were no cases in which the back wall of a vessel became inadvertently caught in the anastomosis. The overall survival rate for distal digital replants was 85% (6/7 replants). It is very difficult to evenly anastomose vessels of differing diameter, especially on a supermicrosurgical scale. In this respect, the IVaS method plays a role in stably anchoring the 2 vessel ends, allowing for the even spacing of suture knots, even in vessels of different caliber. Because of its ease of use and exactitude, many surgeons may be able to use the IVaS method to reliably complete small anastomoses in fingertip replantations.

IFATS collection: Fibroblast growth factor-2-induced hepatocyte growth factor secretion by adipose-derived stromal cells inhibits postinjury fibrogenesis through a c-Jun N-terminal kinase-dependent mechanism.

Adipose-derived stem/stromal cells (ASCs) not only function as tissue-specific progenitor cells but also are multipotent and secrete angiogenic growth factors, such as hepatocyte growth factor (HGF), under certain circumstances. However, the biological role and regulatory mechanism of this secretion have not been well studied. We focused on the role of ASCs in the process of adipose tissue injury and repair and found that among injury-associated growth factors, fibroblast growth factor-2 (FGF-2) strongly promoted ASC proliferation and HGF secretion through a c-Jun N-terminal kinase (JNK) signaling pathway. In a mouse model of ischemia-reperfusion injury of adipose tissue, regenerative changes following necrotic and apoptotic changes were seen for 2 weeks. Acute release of FGF-2 by injured adipose tissue was followed by upregulation of HGF. During the adipose tissue remodeling process, adipose-derived 5-bromo-2-deoxyuridine-positive cells were shown to be ASCs (CD31-CD34+). Inhibition of JNK signaling inhibited the activation of ASCs and delayed the remodeling process. In addition, inhibition of FGF-2 or JNK signaling prevented postinjury upregulation of HGF and led to increased fibrogenesis in the injured adipose tissue. Increased fibrogenesis also followed the administration of a neutralizing antibody against HGF. FGF-2 released from injured tissue acts through a JNK signaling pathway to stimulate ASCs to proliferate and secrete HGF, contributing to the regeneration of adipose tissue and suppression of fibrogenesis after injury. This study revealed a functional role for ASCs in the response to injury and provides new insight into the therapeutic potential of ASCs.

Evaluation of animal models for the hair-inducing capacity of cultured human dermal papilla cells.

BACKGROUND: The dermal papilla (DP) interacts with epithelial cells for folliculogenesis. For translational research on cell therapies for hair regrowth with cultured human DP cells (hDPCs), a model to evaluate the capacity of hDPCs to induce hair formation is inevitable. METHODS: Chamber models were constructed by transplanting 4 different combinations of mouse or human epithelial and mesenchymal cells into a silicone chamber implanted onto the back of nude mice. In parallel, 3 types of sandwich constructs were created by inserting hDPCs or human DP tissue between the epidermis and dermis of isolated rat footpad skin or human facial skin, and subcutaneously transplanting the constructs into the back of nude mice. Four to six weeks later, skin sections of each model were histologically examined. RESULTS: Folliculoneogenesis was detected in both chamber and sandwich models, although the induction rate and maturity of the hair follicles varied among cell combination subgroups in each model. The difference in hair induction rate was not statistically significant between 2 representative chamber and sandwich subgroups using cultured hDPCs. The sandwich model, however, required fewer hDPCs, did not require human keratinocytes, and exhibited a higher rate of successful sample collection. CONCLUSIONS: Although there is no significant difference in hair induction rate, the sandwich model using cultured hDPCs and the rat sole skin is more feasible than the chamber model using human cultured keratinocytes and hDPCs as a tool to evaluate the hair-inducing capacity of cultured hDPCs.

Influences of centrifugation on cells and tissues in liposuction aspirates: optimized centrifugation for lipotransfer and cell isolation.

BACKGROUND: Although injective autologous fat transplantation is one of the most attractive options for soft-tissue augmentation, problems such as unpredictability and fibrosis resulting from fat necrosis limit its universal acceptance. Centrifugation is one of most common methods for overcoming these difficulties. This study was performed to investigate quantitatively the effects of centrifugation on liposuction aspirates to optimize centrifugal conditions for fat transplantation and isolation of adipose-derived stem cells. METHODS: Liposuction aspirates, obtained from eight healthy female donors, were either not centrifuged or centrifuged at 400, 700, 1200, 3000, or 4200 g for 3 minutes. The volumes of the oil, adipose, and fluid portions and numbers of blood cells and adipose-derived cells in each portion were examined. The processed adipose tissues (1 ml) were injected into athymic mice, and grafts were harvested and weighed at 4 weeks. Morphologic alterations were observed using light and scanning electron microscopy. RESULTS: Centrifugation concentrated adipose tissues and adipose-derived stem cells in the adipose portion and partly removed red blood cells from the adipose portion. Centrifugation at more than 3000 g significantly damaged adipose-derived stem cells. Centrifugation enhanced graft take per 1 ml centrifuged adipose but reduced calculated graft take per 1 ml adipose before centrifugation. CONCLUSIONS: Excessive centrifugation can destroy adipocytes and adipose-derived stem cells, but appropriate centrifugation concentrates them, resulting in enhanced graft take. The authors tentatively recommend 1200 g as an optimized centrifugal force for obtaining good short- and long-term results in adipose transplantation.

Intravascular stenting (IVaS) for safe and precise supermicrosurgery.

The diameter of very small vessels (about 0.5 mm or less) causes difficulties in placing forceps into the lumen and in completing anastomosis without inadvertently catching the back wall during supermicrosurgery. The insertion of nylon monofilaments into small vessels has overcome this problem. We implanted superficial inferior epigastric arterial (SIEA) flaps in 10 rats and also performed supermicroanastomosis (diameter, 0.15 mm) using SIEA flaps in mice. The back wall was never inadvertently caught using the intravascular stenting (IVaS) method, and the immediate patency rate was 100%. An advantage of using nylon for IVaS is that various sizes can be selected. We successfully anastomosed vessels with a minimum diameter of 0.15 mm. Even smaller vessels can be precisely and safely anastomosed using the IVaS method.

Preserved proliferative capacity and multipotency of human adipose-derived stem cells after long-term cryopreservation.

BACKGROUND: Human adipose-derived stem (stromal) cells are promising as a regenerative therapy tool for defective tissues of mesenchymal lineage, including fat, bone, and cartilage, and blood vessels. In potential future clinical applications, adipose-derived stem cell cryopreservation could be an indispensable fundamental technology, as has occurred in other fields involving cell-based therapies using hematopoietic stem cells and umbilical cord blood cells. METHODS: The authors examined the proliferative capacity and multipotency of human adipose-derived stem cells isolated from lipoaspirates of 18 patients in total before and after a 6-month cryopreservation following their defined protocol. Proliferative capacity was quantified by measuring doubling time in cell culture, and multipotency was examined with differentiation assays for chondrogenic, osteogenic, and adipogenic lineages. In addition, expression profiles of cell surface markers were determined by flow cytometry and compared between fresh and cryopreserved adipose-derived stem cells. RESULTS: Cryopreserved adipose-derived stem cells fully retained the potential for differentiation into adipocytes, osteoblasts, and chondrocytes and for proliferative capacity. Flow cytometric analyses revealed that surface marker expression profiles remained constant before and after storage. CONCLUSIONS: Adipose-derived stem cells can be cryopreserved at least for up to 6 months under the present protocol without any loss of proliferative or differentiation potential. These results ensure the availability of autologous banked adipose-derived stem cells for clinical applications in the future.

Influences of preservation at various temperatures on liposuction aspirates.

BACKGROUND: Aspirated fat is not only a filler material but also an abundant source of adipose-derived stem cells. The aim of this study was to assess degeneration of aspirated fat during preservation and optimize the preservation method for lipoaspirates. METHODS: Aspirated fat was preserved at room temperature for 1, 2, 4, and 24 hours (n = 10 each); at 4 degrees C for 1, 2, and 3 days (n = 14 each); or at -80 degrees C for 1 month (n = 3). Morphologic changes were assessed with scanning electron microscopy. Adipose-derived stem cell yield was measured after 1 week of culture. For aspirated fat preserved at room temperature, damaged adipocytes were assessed by measuring the oil volume ratio after centrifugation (n = 6) and glycerol-3-phosphate-dehydrogenase activity in washing solution (n = 4). Cell surface marker expression was examined by flow cytometry (n = 3). RESULTS: Although the scanning electron microscopic assay indicated no remarkable anatomical changes based on preservation methods, oil volume significantly increased in fat preserved at room temperature for 4 hours. Adipose-derived stem cell yield was significantly reduced by preservation at room temperature for 24 hours and by preservation at 4 degrees C for 2 or 3 days. Flow cytometric analysis suggested that the biological properties of adipose-derived stem cells did not significantly change at 4 degrees C up to 3 days. The cells were isolated from cryopreserved fat, but the yield was much less than that from fresh aspirated fat. CONCLUSIONS: Aspirated fat should be transplanted as quickly as possible if it is preserved at room temperature. For adipose-derived stem cell isolation, aspirated fat can be stored or transported overnight if it is preserved at 4 degrees C without adipose-derived stem cell yield loss or changes in biological properties.

A clinical trial of topical bleaching treatment with nanoscale tretinoin particles and hydroquinone for hyperpigmented skin lesions.

BACKGROUND: Although combined use of tretinoin (all-trans-retinoic acid; atRA) and hydroquinone improves various hyperpigmented lesions, the pharmacologic instability of atRA and atRA-induced irritant dermatitis are difficult unsolved problems. OBJECTIVE: The objective was to evaluate the efficacy and adverse effects of a newly formulated gel containing inorganic-coated atRA nanoscale particles (nano-atRA gel). METHODS: Nano-atRA gel was used in our two-phased bleaching protocol: 5% hydroquinone and 7% lactic acid ointment were used along with nano-atRA gel in the bleaching phase (2-8 weeks), and 5% hydroquinone and 7% ascorbic acid ointment were used alone during the healing phase (4-8 weeks). Eighty-four patients with facial hyperpigmented lesions were enrolled in this study, and 77 of them (88 lesions) followed up for more than 10 weeks were analyzed. RESULTS: Hyperpigmentation was improved in 84 of 88 lesions (95.5%) after a mean treatment period of 14.3 weeks and was almost eliminated in 52 lesions (59.1%). Nano-atRA gel caused exfoliation and scaling similar to that seen with conventional atRA gel, whereas the erythema seen in the bleaching phase appeared to be weaker. CONCLUSION: Nano-atRA gel can improve hyperpigmentation to a similar extent as conventional atRA gel. It also induces irritant dermatitis, but with less erythema.

Characterization of wound drainage fluids as a source of soluble factors associated with wound healing: comparison with platelet-rich plasma and potential use in cell culture.

Wound fluids, human serum from platelet-poor and platelet-rich plasma (SPPP and SPRP), contain various soluble factors involved in cell growth and proliferation. Levels of cytokines, chemokines, and matrix metalloproteinases (MMPs) in drainage fluids (DFs) harvested from subcutaneous wounds, punctured fluids (PF) from seroma, and SPPP were measured. SPPP and SPRP from four healthy volunteers were also subjected to the analysis. Biochemical profiles of DF reflected the sequential stages of wound healing. Early-phase DF contained high concentrations of basic fibroblast growth factor and platelet-derived growth factor and EGF. The levels of keratinocyte growth factor, interleukin-6, and MMP-8 in DF peaked on days 2-3, while vascular endothelial growth factor, hepatocyte growth factor, interleukin-8, and MMP-1 increased over time during days 0-6. Punctured fluids contained high levels of TGF-beta1, keratinocyte growth factor, vascular endothelial growth factor, hepatocyte growth factor, and MMP-1. Experiments using human adipose-derived stem cells and dermal fibroblasts cultured in media containing various concentrations of DF and fetal bovine serum suggested that for some cell types, DF-contained growth factors that are not obtained from SPRP could be used to supplement or substitute for serum in culture media. SPRP and DF are economical ready-made mixtures of serum and autologous soluble factors, and may be differentially useful for regenerative therapies.