＜Editors' Choice＞ Effects of exosomes derived from the induced pluripotent stem cells on skin wound healing.

Recently, the effects of stem cell supernatants or exosomes, such as skin wounds, have attracted attention. However, the effects of the induced pluripotent stem (iPS) cell-derived exosomes (iPS-Exos) have not been investigated in detail. Here, we investigated the effects of iPS-Exos on skin wound healing using an animal model. We isolated iPS-Exos from the iPS cell culture media. Control exosomes were isolated from unused iPS cell culture media (M-Exos). We first observed the morphologic characteristics of the isolated exosomes and examined the expression of surface antigens. The effects of these exosomes on the migratory response and proliferation of fibroblasts were analyzed as well. Additionally, using a diabetic ulcer model, the effects of iPS-Exos and M-Exos on skin wound healing were investigated. Transmission electron microscope analysis demonstrated that the size of iPS-Exos (120 ± 25 nm) was significantly larger than that of M-Exos (≤ 100 nm). Flow cytometry analyses showed that iPS-Exos were positive for CD9, CD63, and CD81, whereas they were negative for HLA-ABC and -DR expression. The migratory ability of fibroblasts cocultured with iPS-Exos was shown to be higher than that of the cells cocultured with M-Exos, as demonstrated using scratch assay. Skin wound healing model results showed that the administration of iPS-Exos results in a faster wound closure compared with that observed in the M-Exo group. In conclusion, the results obtained here indicate that iPS-Exos may promote the migration of fibroblasts in vitro and in vivo, suggesting the possibility of using iPS-Exos for the treatment of diabetic ulcer.

Effects of Tissue Component Volumes on Vascular Resistance in Free Flaps.

Background A successful free flap transfer is achieved, in part, by having a thorough understanding of vascular anatomy and blood flow dynamics. We previously reported that vascular resistance differs by type of free flap. To test the hypothesis that the difference reflects the proportion of tissue components within free flaps, we calculated blood flow and vascular resistance for free flaps in which we determined the volume of each tissue component. Methods Measurements and calculations were made for 40 free flap transfers performed at our hospital: 7 radial forearm flaps, 14 anterolateral thigh flaps, and 19 rectus abdominis myocutaneous flaps. Results The vascular resistance of free flaps was inversely related to the volume of each tissue component. Univariate regression analysis revealed that muscle volume correlated most closely with resistance (r = 0.881), followed by skin (r = 0.622), and fat (r = 0.577). Multiple regression analysis confirmed the relationship between combined muscle and fat volume and resistance (R2 = 0.865). Conclusions A strong inverse correlation exists between vascular resistance and combined muscle and fat tissue volume in flaps. It may be helpful to consider these relationships when making decisions regarding choice of free flap and recipient vessels.

Skin-derived precursor cells promote wound healing in diabetic mice.

BACKGROUND: Impaired wound healing as one of the complications arising from diabetes mellitus is a serious clinical issue. Recently, various cell therapies have been reported for promotion of wound healing. Skin-derived precursor cells (SKPs) are multipotent adult stem cells with the tendency to differentiate into neurons. We investigated the potency of promoting diabetic wound healing by the application of SKPs. METHODS: Skin-derived precursor cells isolated from diabetic murine skin were cultured in sphere formation medium. At passage 2, they were suspended in phosphate-buffered saline (PBS), and applied topically to full-thickness excisional cutaneous wounds in diabetic mice. Application of PBS served as controls (n = 21 for each group; n = 42 total). Time to closure and percentage closure were calculated by morphometry. Wounds were harvested at 10 and 28 days and then processed, sectioned, and stained (CD31, α-smooth muscle actin, and neurofilament heavy chain) to quantify vascularity and neurofilaments. RESULTS: Wounds treated with SKPs demonstrated a significantly decreased time to closure (18.63 days) compared with PBS-control wounds (21.72 days, P < 0.01), and a significant improvement in percentage closure at 7, 10, 14, and 18 days compared with PBS-control wounds (P < 0.01). Histological analysis showed that the Capillary Score (the number of vessels/mm2) was significantly higher in SKP-treated wounds at day 10 but not at day 28. Nerve Density (the number of neurofilaments/mm2) had increased significantly in SKP-treated wounds at day 28 compared with control group. Some applied SKPs were stained by neurofilament heavy chain, which demonstrates that SKPs directly differentiated into neurons. CONCLUSIONS: Skin-derived precursor cells promoted diabetic wound healings through vasculogenesis at the early stage of wound healing. Skin-derived precursor cells are a possible therapeutic tool for diabetic impaired wound healing.

Head and neck free flap reconstruction under the COVID-19 pandemic.

High perioperative mortality and complication rates during the coronavirus disease 2019 (COVID-19) pandemic have been reported. In head and neck reconstruction, not only is patient safety important, but the prevention of infection introduced by the surgical team is also important because the procedure is performed in close proximity to the upper respiratory tract. In addition, recent studies have reported an increased risk for thrombus formation after infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) or COVID-19 vaccination, which is problematic for microsurgical reconstruction procedures. At the authors' institution, patients undergoing head and neck reconstruction are requested to stay home for 2 weeks and undergo screening tests for COVID-19 before admission. Surgeons use standard personal protective equipment during surgery. There was no significant difference in the rate of total flap necrosis between the COVID-19 and non-pandemic periods or large difference of perioperative complication rates between vaccinated and non-vaccinated patients. No surgery-related infections among the surgical staff were also found.

Exclusive expression of KANK4 promotes myofibroblast mobility in keloid tissues.

Keloids are characterized by abnormal wound healing with excessive accumulation of extracellular matrix. Myofibroblasts are the primary contributor to extracellular matrix secretion, playing an essential role in the wound healing process. However, the differences between myofibroblasts involved in keloid formation and normal wound healing remain unclear. To identify the specific characteristics of keloid myofibroblasts, we initially assessed the expression levels of well-established myofibroblast markers, α-smooth muscle actin (α-SMA) and transgelin (TAGLN), in scar and keloid tissues (n = 63 and 51, respectively). Although myofibroblasts were present in significant quantities in keloids and immature scars, they were absent in mature scars. Next, we conducted RNA sequencing using myofibroblast-rich areas from keloids and immature scars to investigate the difference in RNA expression profiles among myofibroblasts. Among significantly upregulated 112 genes, KN motif and ankyrin repeat domains 4 (KANK4) was identified as a specifically upregulated gene in keloids. Immunohistochemical analysis showed that KANK4 protein was expressed in myofibroblasts in keloid tissues; however, it was not expressed in any myofibroblasts in immature scar tissues. Overexpression of KANK4 enhanced cell mobility in keloid myofibroblasts. Our results suggest that the KANK4-mediated increase in myofibroblast mobility contributes to keloid pathogenesis.

＜Editors' Choice＞ Supernatant from activated omentum accelerates wound healing in diabetic mice wound model.

Diabetic wounds are considered one of the most frequent and severe complications of diabetes mellitus. Recently, the omentum has been used in diabetic wound healing because of its tissue repair properties. The activated omentum is richer in growth factors than the inactivated, thereby contributing to the wound healing process. To further investigate the effect of activated omentum conditioned medium (aOCM) on diabetic wound healing, we injected supernatant from aOCM, saline-OCM (sOCM), inactivated-OCM (iOCM), and medium (M) subcutaneously upon creation of a cutaneous wound healing model in diabetic mice. Wound area (%) was evaluated on days 0, 3, 5, 7, 9, 11, 14, 21, and 28 post-operation. At 9 and 28 d post-operation, skin tissue was harvested and assessed for gross observation, neovascularization, peripheral nerve fiber regeneration, and collagen deposition. We observed that aOCM enhanced the wound repair process, with significant acceleration of epidermal and collagen deposition in the surgical lesion on day 9. Additionally, aOCM displayed marked efficiency in neovascularization and peripheral nerve regeneration during wound healing. Thus, aOCM administration exerts a positive influence on the diabetic mouse model, which can be employed as a new therapy for diabetic wounds.

Artificial PGA/Collagen-based Bilayer Conduit in Short Gap Interposition Setting Provides Comparable Regenerative Potential to Direct Suture.

The aim of this study was to evaluate whether the Nerbridge, an artificial polyglycolic acid conduit with collagen matrix, is comparable to direct nerve suture in a rat sciatic nerve injury model in a short-gap interposition (SGI) setting. Methods: Sixty-six female Lewis rats were randomly divided into the sham group (n = 13); no reconstruction (no-recon) group (n = 13; rat model with 10 mm sciatic nerve defect); direct group (n = 20; rat sciatic nerve injury directly connected by 10-0 Nylon); and SGI group (n = 20; sciatic nerve injury repaired using 5-mm Nerbridge). Motor function and histological recovery were evaluated. The sciatic nerve and gastrocnemius muscle were harvested for quantification of the degree of nerve regeneration and muscle atrophy. Results: The SGI and direct groups achieved equal recovery in both functional and histological outcomes. At weeks 3 and 8 postsurgery, there was a significant improvement in the sciatic functional index of the SGI group when compared with that of the no-recon group (P < 0.05). Furthermore, the direct and SGI groups had less muscle atrophy at 4 and 8 weeks postsurgery compared with the no-recon group (P < 0.05). The axon density and diameter at the distal site in the SGI group were significantly higher than that in the no-recon group and comparable to that in the direct and sham groups. Conclusion: An artificial nerve conduit has equal potential as direct suture in motor nerve reconstruction when used in the SGI setting.

Expansion Culture of Hair Follicle Stem Cells through Uniform Aggregation in Microwell Array Devices.

Hair regeneration using hair follicle stem cells (HFSCs) and dermal papilla cells is a promising approach for the treatment of alopecia. One of the challenges faced in this approach is the quantitative expansion of HFSCs while maintaining their hair induction capacity. In this study, HFSC expansion was achieved through the formation of uniform-diameter cell aggregates that were subsequently encapsulated in Matrigel. We designed a microwell array device, wherein mouse HFSCs were seeded, allowed to form loosely packed aggregates for an hour, and then embedded in Matrigel. Quantitative analysis revealed a 20-fold increase in HFSC number in 2 weeks through this culture device. Gene expression of trichogenic stem cell markers in the device-grown cells showed a significant increase compared with that of typical flat substrate Matrigel suspension culture cells. These microwell array-cultured HFSCs mixed with freshly isolated embryonic mesenchymal cells indicated vigorous hair regeneration on the skin of nude mice. Furthermore, we examined the feasibility of this approach for the expansion of human HFSCs from androgenetic alopecia patients and found that the ratio of CD200+ cells was improved significantly in comparison with that of cells cultured in a typical culture dish or in a Matrigel suspension culture on a flat substrate. Therefore, the novel approach proposed in this study may be useful for HFSC expansion in hair regenerative medicine.

＜Editors' Choice＞ Long-term outcomes of lateral skull base reconstruction with a free omental flap and facial nerve reconstruction.

In lateral skull base reconstruction, it is necessary to seal the defect in the lateral skull base, fill the dead space, and, sometimes, reconstruct the facial nerve. However, this procedure is difficult to perform with a standard musculocutaneous flap. Therefore, for such cases, an omental flap is used in our hospital because of its flexibility. In this study, we report our experience with the procedure (lateral skull base reconstruction with a free omental flap) and its long-term outcome and facial nerve reconstruction, with special focus on facial nerve recovery. This study is a technical note and a retrospective review. It was conducted in Nagoya University Hospital. Overall, 16 patients (12 women and 4 men; mean age: 55.1 years) underwent lateral skull base reconstruction with a free omental flap after subtotal temporal bone resection or lateral temporal bone resection during 2005-2017. The main outcome measures were postoperative complications and facial nerve recovery: Yanagihara score and House-Brackmann grading system. Complications included partial necrosis and minor cerebrospinal fluid leakage in 2 patients. Facial nerve recovery could be observed more than 12 months after surgery, with a mean Yanagihara score of 19.6 and House-Brackmann grade of 3.60. The free omental flap is a reliable method for lateral skull base reconstruction, especially in cases where facial nerve reconstruction is needed. To the best of our knowledge, this is the first report on facial nerve recovery after lateral skull base reconstruction.

Development of a Novel Scar Screening System with Machine Learning.

BACKGROUND: Hypertrophic scars and keloids tend to cause serious functional and cosmetic impediments to patients. As these scars are not life threatening, many patients do not seek proper treatment. Thus, educating physicians and patients regarding these scars is important. The authors aimed to develop an algorithm for a scar screening system and compare the accuracy of the system with that of physicians. This algorithm was designed to involve health care providers and patients. METHODS: Digital images were obtained from Google Images (Google LLC, Mountain View, Calif.), open access repositories, and patients in the authors' hospital. After preprocessing, 3768 images were uploaded to the Google Cloud AutoML Vision platform and labeled with one of the four diagnoses: immature scars, mature scars, hypertrophic scars, and keloid. A consensus label for each image was compared with the label provided by physicians. RESULTS: For all diagnoses, the average precision (positive predictive value) of the algorithm was 80.7 percent, the average recall (sensitivity) was 71 percent, and the area under the curve was 0.846. The algorithm afforded 77 correct diagnoses with an accuracy of 77 percent. Conversely, the average physician accuracy was 68.7 percent. The Cohen kappa coefficient of the algorithm was 0.69, while that of the physicians was 0.59. CONCLUSIONS: The authors developed a computer vision algorithm that can diagnose four scar types using automated machine learning. Future iterations of this algorithm, with more comprehensive accuracy, can be embedded in telehealth and digital imaging platforms used by patients and primary doctors. The scar screening system with machine learning may be a valuable support tool for physicians and patients. CLINICAL QUESTION/LEVEL OF EVIDENCE: Diagnostic, II.

Stem cells from human exfoliated deciduous teeth (SHED) enhance wound healing and the possibility of novel cell therapy.

BACKGROUND AIMS: In recent years, stem cells from human exfoliated deciduous teeth (SHED) have received attention as a novel stem cell source with multipotent potential. We examined the effect on wound-healing promotion with unique stem cells from deciduous teeth as a medical waste. METHODS: An excisional wound-splinting mouse model was used and the effect of wound healing among SHED, human mesenchymal stromal cells (hMSCs), human fibroblasts (hFibro) and a control (phosphate-buffered saline; PBS) was evaluated by macroscopy, histology and enzyme-linked immunosorbent assay (ELISA), and the expression of hyaluronan (HA), which is related to wound healing, investigated. RESULTS: SHED and hMSCs accelerated wound healing compared with hFibro and the control. There was a statistically significant difference in wound healing area among hFibro, hMSCs and SHED compared with the control after day 5. At days 7 and 14 after cell transplantation, the histologic observation showed that transplanted PKH26-positive cells were surrounded by human HA binding protein, especially in hMSCs and SHED. HA expression volume values were 1558.41 ± 60.33 (control), 2092.75 ± 42.56 (hFibro), 2342.07 ± 188.10 (hMSCs) and 2314.85 ± 164.91 (SHED) ng/mg, respectively, and significantly higher in hMSCs and SHED compared with hFibro and control at days 7 and 14 (P < 0.05). CONCLUSIONS: Our results show that SHED hMSCs have similar effects of wound-healing promotion as hFibro and controls. This implies that SHED might offer a unique stem cell resource and the possibility of novel cell therapies for wound healing in the future.

Human deciduous teeth dental pulp cells with basic fibroblast growth factor enhance wound healing of skin defect.

In this research, we examined the effect on wound healing applying basic fibroblast growth factor (b-FGF) that is approved for clinical use to enhance wound healing and human deciduous teeth dental pulp cells (hDPCs) in clinics, but that have been attracting attention as a novel stem cell source in recent years. Human deciduous teeth were harvested from healthy volunteers, and hDPCs were isolated. We used a nude mouse full-thickness skin defect model and evaluated wound healing by macroscopic view and histologic and histomorphometric analysis. The mice were randomly divided into 4 groups: phosphate-buffered saline-treated group (control group), b-FGF-treated group (b-FGF group), hDPC-treated group (hDPC group), and hDPC and b-FGF-treated group (hDPC/b-FGF group). Basic fibroblast growth factor and hDPC groups accelerated wound healing compared with the control group. There was no statistically significant difference in wound healing observed between the hDPC and b-FGF groups. The hDPC/b-FGF group demonstrated accelerated wound healing compared with other groups. At day 14, PKH26-positive cells were surrounded by human type I collagen in hDPC and hDPC/b-FGF groups in immunohistologic evaluation. Significantly increased collagen fibril areas in wound tissues were observed in b-FGF, hDPC, and hDPC/b-FGF groups as compared with the control group at days 7 and 14. Our results showed that the hDPC/b-FGF group significantly promotes wound healing compared with other groups. This study implies that deciduous teeth that are currently considered as medical spare parts might offer a unique stem cell resource for potential of new cell therapies for wound healing in combination with b-FGF.

Peripheral Nerve Regeneration Using Different Germ Layer-Derived Adult Stem Cells in the Past Decade.

Peripheral nerve injuries (PNIs) are some of the most common types of traumatic lesions affecting the nervous system. Although the peripheral nervous system has a higher regenerative ability than the central nervous system, delayed treatment is associated with disturbances in both distal sensory and functional abilities. Over the past decades, adult stem cell-based therapies for peripheral nerve injuries have drawn attention from researchers. This is because various stem cells can promote regeneration after peripheral nerve injuries by differentiating into neural-line cells, secreting various neurotrophic factors, and regulating the activity of in situ Schwann cells (SCs). This article reviewed research from the past 10 years on the role of stem cells in the repair of PNIs. We concluded that adult stem cell-based therapies promote the regeneration of PNI in various ways.

A Novel Technique to Lengthen the Reverse Latissimus Dorsi Muscle Flap Arc.

Reconstruction of the lower lumbar region is challenging for surgeons due to limited locoregional flap choices. The latissimus dorsi muscle flap is a mainstay for this area; however, there are several limitations, including that the dominant thoracodorsal artery and vein pedicle-based flaps are not reachable for reconstruction of the lumbar region, while perforator of intercostal artery and veins pedicle-based reverse latissimus dorsi (RLD) flap mobility is limited by including multiple perforators. Here, we describe a novel operative technique that lengthens the rotation arc of RLD muscle flaps. The surgical technique is as follows: RLD is elevated based on lower perforator of intercostal artery and veins (usually including two of the eighth-11th perforators); thoracodorsal artery and vein are ligated; and the flap is mobilized toward the defect. When RLD was not reachable to the defect, the far aspect of the intercostal artery and vein from the defect was ligated and the perforator was elevated with the near aspect of the intercostal artery and vein from intercostal space. Because the intercostal space measured between approximately 3 cm and 4 cm, this dissection gained 3-4 cm of rotational arc per intercostal space. Moreover, because the lower ribs follow a medio-cranial to latero-caudal direction, this dissection enabled the flap to extend latero-caudally or medio-cranially while maintaining its blood supply. Other applications using this technique may involve expanding the RLD flap arc caudally, ventrally, and ipsilaterally. We believe this new technique provides a reliable alternative for lower back reconstruction.

Repair of temporal branch of the facial nerve with novel polyglycolic acid-collagen tube: a case report of two cases.

Autologous nerve transplantation has been the gold standard in the treatment of facial nerve injury, however it has not been achieved satisfactory result and needs donor sacrifice. A polyglycolic acid collagen conduit (Nerbridge, Toyobo Co., Japan) has the potential to compare to or exceed autologous nerve grafts in promoting nerve regeneration. Here we report two cases of traumatic temporal facial nerve injury repairs with Nerbridge. The severed temporal branch of the facial nerve was repaired with Nerbridge conduits in two patients. Recovery of movement was assessed by clinical photography and needle electromyography. The frontal muscle started moving five months postoperatively in both cases. Electromyography at twelve months showed polymorphic electric discharge, suggesting connection of the injured nerve to the frontal muscle. In the final results, each patient had good eyebrow elevation distance and moderate forward gaze recovery in comparison to their healthy sides. Considering that facial nerves are reported to recover incompletely even in autologous nerve graft repair cases, our two cases showed reasonable recovery comparable to nerve autografting. The Nerbridge conduit is a promising alternative to standard treatments for facial nerve recovery.

Multiple Thrombi during Microvascular Anastomosis Caused by Decreased Antithrombin Activity: A Case Report.

With recent advances in microsurgical instruments and technique, microvascular anastomosis has become a universal surgical technique; however, thrombosis still presents in a number of cases. Tension, twisting, and compression to the anastomotic site are the main causes of thrombus; however, disorder of the coagulation-fibrinolysis system also need to be considered. To date, only few reports exist regarding thrombosis caused by disorder of coagulant system in microvascular anastomosis. Here we report our 3 cases in which multiple thrombus formation occurred intraoperatively caused by decrease of antithrombin (AT) activity. AT activity was measured twice a day after vascular anastomosis: after surgery and up to 3 days after surgery. Thrombosis was not observed in any of the 3 patients intraoperatively after the transfusion, or thrombosis was not observed in any of 3 patients intraoperatively after the transfusion or postoperatively, and no other complications were observed. In these 3 cases, the thrombus was not caused by technical error or other previously described factors. The observed intraoperative decrease in AT activity was thought to be caused by thrombus formation. It is important that microsurgeons are reminded that disorders of the coagulation-fibrinolysis system could cause thrombosis.

Correlation between blood flow, tissue volume and microvessel density in the flap.

The purpose of this study was to assess the correlation between tissue volume and blood flow of the flap in an animal model. Using animal model, tissue volume can be attenuated, and precise change of blood flow could be evaluated. We further investigate the relationship between blood flow and vascular density in the tissue. In this study, we assessed flap conductance (ml/min/mm Hg) as to evaluate the conductivity of blood flow into the flap. Japanese white rabbit was used (n = 7) for this study. The amount of blood flow of jejunal and latissimus dorsi muscle (LD) flaps was measured while removing the distal portion of the flap sequentially. Conductance at each time was calculated from blood pressure and blood flow volume. The tissue volume at each time was also measured. The correlation between conductance and volume was analyzed using a linear mixed model. Immunohistochemical evaluation of microvessel densities (MVD) in these tissues was also performed for CD31/PECAM1 positive area. Conductance and tissue volume were significantly correlated in both jejunal and LD flaps. As the volume increases by 1 cm3, the conductance increased significantly by 0.012 ml/min/mm Hg in jejunum, and by 0.0047 ml/min/mm Hg in LD. Mean MVD was 1.15 ± 0.52% in the jejunum and 0.37 ± 0.29% in the LD muscle. In this study, we revealed that flap conductance is proportional to volume and proportional constant is different between the type of tissue. It suggests that the difference of MVD creates the unique conductance of each tissue.

Erratum: Increased Defect Size is Associated with Increased Complication Rate after Free Tissue Transfer for Midanterior Skull-Base Reconstruction.

[This corrects the article DOI: 10.1055/s-0038-1676777.].

Increased Defect Size is Associated with Increased Complication Rate after Free Tissue Transfer for Midanterior Skull-Base Reconstruction.

Objective Complications after skull-base reconstruction are often problematic. We consider that local factors, for example, localization of defect areas are possible risk factors. This study aimed to investigate our case series of skull-base reconstructions in our institution and to identify local risk factors that predispose to wound complications. Design This study is presented as a retrospective study. Setting Research work was took place at Nagoya University Hospital. Participants Forty-eight patients who had undergone reconstruction after midanterior skull-base resection between January 2004 and December 2015 were included in this study. Defects apart from the skull-base were categorized into nasal and paranasal cavity (N), orbit (O), palate (P), and facial skin (S). Postoperative local complications including cerebrospinal fluid (CSF) leakage, local infection, wound dehiscence (fistula in face or palate), and flap necrosis (partial or total) were investigated. Main Outcome Measures Main outcome measures were postoperative complications in patients with each defect. Results Apart from the skull-base, defects included 28 ONP (58.3%), 10 ONPS (20.8%), 3 ON (6.3%), 3 ONS (6.3%), 1 NP (2.1%), and 1 OS (2.1%). Comparison based on numbers of resected regions revealed that a significantly higher complication rate was seen in patients with four resected regions than in those with three regions (90.0% vs. 45.2%, p < 0.05). Conclusion There was a trend suggesting that more resected regions corresponded to a greater risk of complications in midanterior skull-base reconstruction. Reconstructive surgeons need to carefully consider the reconstruction of such complicated defects.

[Regenerative medicine for anti-aging].

Regenerative medicine refers to research and treatments that restore damaged tissues. It consists of three factors: cells, chemical substances, and scaffolds. One of its purposes is rejuvenation, and from this point of view, it really resembles anti-aging medicine. Recently, studies concerning stem cells such as embryonic stem cells (ES) and induce pluripotent stem cells (iPS) have made remarkable progress and seem promising, but are still far from clinical use. In contrast, we introduce a few examples of anti-aging medicine using regenerative medicine methods that have been applied successfully in clinical use such as cultured fibroblast, adipose tissue derived stem cells and platelet rich plasma (PRP), and continue researching cultured dermal papilla cells.