The effects of photodynamic therapy with blue light and papain-based gel associated with Urucum, on collagen and fibroblasts: a spectroscopic and cytotoxicity analysis.

Papacarie Duo™ is clinically used and has proven effectiveness; however, it is necessary to improve its antimicrobial action. The combined treatment of Papacarie Duo™ with Urucum (Bixa Orellana) could create a potential tool for dental caries treatment; its extract obtained from the seeds' pericarp contains a water-soluble primary pigment (cis-bixin) with smaller amounts of other carotenoids. The dicarboxylic acid salts of cis-norbixin and trans-norbixin occur in heated alkaline solutions. To analyze the absorption spectra and cytotoxicity (with human dermal fibroblasts) in different concentrations of Urucum, associated or not with Papacarie Duo™, we performed this in vitro study. The effects of pure Urucum, Papacarie Duo™, and PapaUrucum™ on the microstructure of collagen were also analyzed. The application of papain-based gel with Urucum did not present cytotoxicity, its exhibit UV absorption spectrum peak around 460 ± 20 nm. Also, it showed that the compound used did not alter the chemical structure of collagen. Consequently, this product could be used as a chemomechanical method to remove dentin caries as well as being a potential product for antimicrobial photodynamic therapy (aPDT) application.

Antimicrobial photodynamic therapy as a new approach for the treatment of vulvovaginal candidiasis: preliminary results.

In this work, we present the efficacy of photodynamic therapy against yeast cells in an animal model. We tested two photosensitizers, methylene blue and protoporphyrin IX. Thirty-seven female BALB-c mice with a body mass of 20-25 g were used. To achieve persistent vaginitis, estrus was induced by subcutaneous injection of 0.1 mg/mL estradiol valerate applied weekly. Three days after pseudo-estrus, intravaginal inoculation with Candida albicans was performed. Mice were anesthetized with ketamine (80 mg/kg) and xylazine (10 mg/kg) by intraperitoneal injection before inoculation, and antimicrobial photodynamic therapy (aPDT) was performed 5 days after fungal inoculation. Two photosensitizers were tested, methylene blue (MB; 100 µM) and protoporphyrin IX (PpNetNI; 10 µM). Two custom-made LEDs emitting light at 660 and 630 nm at approximately 800 mW each were used for irradiation. The aPDT treatment reduced the fungal colony-forming units (CFUs) by one order of magnitude for the MB (p = 0.020) and PpNetNI (p = 0.018) photosensitizers. Seven days after the treatment, there were significantly fewer CFUs compared to the control group (p = 0.041 and p = 0.035 for MB and PpNetNI, respectively), but this was not increased compared to the initial number immediately after aPDT. Using aPDT as a therapeutic option to decrease fungal infection in a vaginal candidiasis model resulted in a significant reduction in the C. albicans population. Both photosensitizers were effective for preventing reinfection within 7 days. The aPDT also had no effect on the vaginal mucosa at the ultrastructural level. In addition to the fungicide effect, we observed reduced swelling and lack of the formation of abscesses, microabscesses coating the cornified epithelial layer, and the accumulation of neutrophils in the submucosa.

Orofacial evaluation in patients with multiple sclerosis using Nordic Orofacial Test-Screening.

OBJECTIVES: The aim of this study is to investigate whether patients with multiple sclerosis (MS) have more orofacial dysfunctions than the general population, using the Nordic Orofacial Test-Screening (NOT-S). MATERIALS AND METHODS: The NOT-S instrument was applied in 34 patients with MS, who went to the MS Reference Center, Universidade Metropolitana de Santos and 34 healthy patients, matched for gender and age. NOT-S results were compared between patients with MS and control subjects. Disability and disease duration were assessed among the patients, in order to establish whether these parameters might affect the results from NOT-S. RESULTS: There was no significant difference in orofacial function between patients with MS and control subjects. There was no statistically significant correlation between disability and NOT-S or between disease duration and NOT-S. However, the correlation between disease duration and the degree of disability was statistically significant, thus suggesting that the results are in accordance with what would be expected regarding MS. CONCLUSIONS: These results indicate that there was no correlation between orofacial dysfunction and MS, although there were some differences in the affected domains. CLINICAL RELEVANCE: This study points out the orofacial dysfunctions which health professionals should be aware in this population.

Increase in cholinergic modulation with pyridostigmine induces anti-inflammatory cell recruitment soon after acute myocardial infarction in rats.

We tested the hypothesis that an increase in the anti-inflammatory cholinergic pathway, when induced by pyridostigmine (PY), may modulate subtypes of lymphocytes (CD4+, CD8+, FOXP3+) and macrophages (M1/M2) soon after myocardial infarction (MI) in rats. Wistar rats, randomly allocated to receive PY (40 mg·kg(-1)·day(-1)) in drinking water or to stay without treatment, were followed for 4 days and then were subjected to ligation of the left coronary artery. The groups-denominated as the pyridostigmine-treated infarcted (IP) and infarcted control (I) groups-were submitted to euthanasia 3 days after MI; the heart was removed for immunohistochemistry, and the peripheral blood and spleen were collected for flow cytometry analysis. Noninfarcted and untreated rats were used as controls (C Group). Echocardiographic measurements were registered on the second day after MI, and heart rate variability was measured on the third day after MI. The infarcted groups had similar MI areas, degrees of systolic dysfunction, blood pressures, and heart rates. Compared with the I Group, the IP Group showed a significant higher parasympathetic modulation and a lower sympathetic modulation, which were associated with a small, but significant, increase in diastolic function. The IP Group showed a significant increase in M2 macrophages and FOXP3(+)cells in the infarcted and peri-infarcted areas, a significantly higher frequency of circulating Treg cells (CD4(+)CD25(+)FOXP3(+)), and a less extreme decrease in conventional T cells (CD25(+)FOXP3(-)) compared with the I Group. Therefore, increasing cholinergic modulation with PY induces greater anti-inflammatory cell recruitment soon after MY in rats.

Effect of laser therapy on immune cells infiltrate after excisional wounds in diabetic rats.

BACKGROUND AND OBJECTIVE: Diabetes alters innate and specific immunity, causing an imbalanced tissue repair process. Very active neutrophils and macrophages are found for a long time in chronic wounds in those individuals. The aim of this study was to evaluate the response of the main effector cells of immunity (neutrophils, macrophages, and T lymphocytes) and to compare the effects of two laser therapy regimens in the postoperative treatment of excision wounds. METHOD: Diabetes was induced in female Wistar rats and a punch was used to cause wounds in the dorsum of each individual. The animals were randomly allocated to a control group (CG), in which the wound was untreated, a single-dose laser group (SLG), in which the wound was submitted to single dose of laser therapy at wavelength of 660 nm, output power of 30 mW, energy density of 4 J/cm(2), and 26-second exposure time, and a fractionated-dose laser group (FLG), submitted to 1 J/cm(2) of laser therapy on Days 1, 3, 8, and 10. Euthanasia was performed on five animals from each group Days 1, 3, 8, 10, 15, and 22. The wound was removed and routinely processed for immunohistochemistry against elastase, CD3, CD68, and CD206 antibodies. The samples were photographed and labeled cells were counted by a blinded observer. The Kruskal-Wallis test was used for the statistical analysis. RESULTS: Neutrophils were predominant in the SLG on Day 1, whereas these cells were mostly found in the CG on Day 3 (P < 0.05). The T lymphocyte count was similar in all groups in the throughout the experiment. On Day 3, the SLG exhibited a greater number of total macrophages (CD68+) (P < 0.05), whereas the macrophage count was similar among the different groups on the other evaluation days. The CD206 + cell counts revealed that the SLG had more M2 macrophages than the CG on Day 8 (P < 0.05), whereas the FLG exhibited more M2 macrophages than the CG on Day 10 (P < 0.05). CONCLUSION: The present findings demonstrate that laser therapy can alter the composition of inflammatory infiltrate in diabetic wounds, leading to a more balanced response transiting from a rapid neutrophil infiltration through to M2 macrophage polarization, especially with a single application of 4 J/cm(2) in the immediate postoperative period.

Immediate results of photodynamic therapy for the treatment of halitosis in adolescents: a randomized, controlled, clinical trial.

UNLABELLED: Light with or without chemical agents has been used to induce therapeutic and antimicrobial effects. With photodynamic therapy, the antimicrobial effect is confined to areas covered by a photosensitive dye and irradiated with light. The aim of the present study was to evaluate the effect of photodynamic therapy for the treatment of halitosis in adolescents through the analysis of volatile sulfur compounds, especially sulfide. A controlled, clinical trial was conducted with 45 adolescents randomly allocated to three groups: group 1, photodynamic therapy administered to the dorsum of the tongue; group 2, treatment with a tongue scraper; and group 3, treatment with a tongue scraper combined with photodynamic therapy. The diagnosis of halitosis was performed using gas chromatography before and after treatment. Comparisons were made using the Kruskal-Wallis test followed by the Student-Newman-Keuls test, with the level of significance set at 5 % (p < 0.05). After treatment, a statistically significant reduction in halitosis was found in all groups (p < 0.001). The greatest reduction in total sulfides (median = 0) occurred with the combination of tongue scraper and photodynamic therapy. The present study describes a novel option for the treatment of halitosis in adolescents with an immediate effect that does not involve the mechanical aggression of the lingual papillae that occurs with conventional treatment. TRIAL REGISTRATION: Photodynamic Therapy in Adolescents Halitosis ( https://clinicaltrials.gov/ct2/show/NCT02007993?term=NCT02007993&rank=1 )Number: NCT02007993FUNDING:FAPESPNumber: 2013/13032-8.

The effect of low-level laser therapy (LLLT) applied prior to muscle injury.

AIM: To evaluate the effect of LLLT (780 nm; 10 J/cm2 , 40 mW, 3.2 J) prior to injury on the morphological analysis, collagen deposition, and activity of matrix metalloproteinase-2 (MMP-2). METHODS: Wistar rats were divided into groups: control; sham; only LLLT; only muscle injury and LLLT + injury. The rats were euthanized at 1, 3, and 7 days following cryoinjury to muscle that was removed for analysis. RESULTS: LLLT applied prior to muscle injury led to a reduction in myonecrosis and inflammatory cells, an increase of blood vessels and immature muscle fibers. An increase in MMP-2 activity and a decrease in collagen deposition were also found, with a better collagen organization and distribution. CONCLUSION: LLLT applied immediately prior to injury had positive effects during the muscle regeneration process. Therefore, this resource may have considerable therapeutic value, especially for athletes who practice sports in which there is a constant risk of muscle injury. Lasers Surg. Med. 47:571-578, 2015. © 2015 Wiley Periodicals, Inc.

Pre-exercise low-level laser therapy improves performance and levels of oxidative stress markers in mdx mice subjected to muscle fatigue by high-intensity exercise.

This study was designed to determine if the levels of oxidative stress markers are influenced by low-level laser therapy (LLLT) in mdx mice subjected to high-intensity exercise training on an electric treadmill. We used 21 C57BL/10ScSn-Dmdmdx/J mice and 7 C57BL/10ScSn mice, all aged 4 weeks. The mice were divided into four groups: a positive control group of normal, wild-type mice (WT); a negative control group of untreated mdx mice; a group of mdx mice that underwent forced high-intensity exercise on a treadmill (mdx fatigue); and another group of mdx mice with the same characteristics that were treated with LLLT at a single point on the gastrocnemius muscle of the hind paw and underwent forced high-intensity exercise on a treadmill. The mdx mice treated with LLLT showed significantly lower levels of creatine kinase (CK) and oxidative stress than mdx mice that underwent forced high-intensity exercise on a treadmill. The activities of the antioxidant enzyme superoxide dismutase (SOD) were higher in control mdx mice than in WT mice. LLLT also significantly reduced the level of this marker. LLLT had a beneficial effect also on the skeletal muscle performance of mdx mice. However, the single application of LLLT and the dose parameters used in this study were not able to change the morphology of a dystrophic muscle.

Facial reconstruction surgery 10 years after treatment for hemangiopericytoma: planning considerations and clinical outcomes.

This paper describes the surgical correction of ankylosis of the right temporomandibular joint and reconstruction of the middle and lower thirds of the right side of the face in a young man who had undergone treatment for hemangiopericytoma 10 years earlier, which led to serious functional, esthetic, and psychological problems and diminished his quality of life. The results of reconstruction surgery with alloplastic material (Medopor®) and surgical planning with fibrotic tissue due to radiation and the risk of osteoradionecrosis are discussed.

Production of extracellular matrix proteins by human pulp fibroblasts in contact with papacárie and carisolv.

PURPOSE: To evaluate the effect of two products for the chemomecanical removal of carious tissue (Papacárie and Carisolv) on human dental pulp fibroblasts and the synthesis of extracellular matrix proteins. MATERIALS AND METHODS: Fibroblasts were divided into three groups: group 1 (control), group 2 (Papacárie) and group 3 (Carisolv). Collagen I, III , fibronectin and osteonectin were analysed by immunofluorescence and compared among the groups. RESULTS: The groups exhibited similar immunolabeling for vimentin, type I collagen and fibronectin, but were negative for type III collagen. Osteonectin staining was strongly positive in the cells treated with Papacárie and Carisolv and weakly positive in the control group. CONCLUSION: The findings of the present study showed that Papacárie and Carisolv are not cytotoxic to pulp fibroblast cells. Moreover, these products stimulate fibroblasts to produce osteonectin, likely leading to the formation of dentin matrix. These findings confirm the safe, beneficial use of both gels in minimally invasive techniques.

Engineering models of head and neck and oral cancers on-a-chip.

Head and neck cancers (HNCs) rank as the sixth most common cancer globally and result in over 450 000 deaths annually. Despite considerable advancements in diagnostics and treatment, the 5-year survival rate for most types of HNCs remains below 50%. Poor prognoses are often attributed to tumor heterogeneity, drug resistance, and immunosuppression. These characteristics are difficult to replicate using in vitro or in vivo models, culminating in few effective approaches for early detection and therapeutic drug development. Organs-on-a-chip offer a promising avenue for studying HNCs, serving as microphysiological models that closely recapitulate the complexities of biological tissues within highly controllable microfluidic platforms. Such systems have gained interest as advanced experimental tools to investigate human pathophysiology and assess therapeutic efficacy, providing a deeper understanding of cancer pathophysiology. This review outlines current challenges and opportunities in replicating HNCs within microphysiological systems, focusing on mimicking the soft, glandular, and hard tissues of the head and neck. We further delve into the major applications of organ-on-a-chip models for HNCs, including fundamental research, drug discovery, translational approaches, and personalized medicine. This review emphasizes the integration of organs-on-a-chip into the repertoire of biological model systems available to researchers. This integration enables the exploration of unique aspects of HNCs, thereby accelerating discoveries with the potential to improve outcomes for HNC patients.

Fully automated algorithm for wound surface area assessment.

Worldwide, clinicians, dentists, nurses, researchers, and other health professionals need to monitor the wound healing progress and to quantify the rate of wound closure. The aim of this study is to demonstrate, step by step, a fully automated numerical method to estimate the size of the wound and the percentage damaged relative to the body surface area (BSA) in images, without the requirement for human intervention. We included the formula for BSA in rats in the algorithm. The methodology was validated in experimental wounds and human ulcers and was compared with the analysis of an experienced pathologist, with good agreement. Therefore, this algorithm is suitable for experimental wounds and burns and human ulcers, as they have a high contrast with adjacent normal skin.

The influence of red laser irradiation timeline on burn healing in rats.

Low-level laser therapy (LLLT) promotes biomodulation of wound healing and literature reports that light delivery during the inflammation could play a different role compared with latter phases of the healing process. The objective of this study was to investigate whether single dose of a red laser (λ = 660 nm) is different from fractionated delivery protocol in full thickness burns. Two lesions were inflicted on the back of 36 rats. In the fractionated dose group (FG), the lesions were irradiated with 1 J/cm² on days 1, 3, 8, and 10 post-wounding. In the single dose group (SG), the lesions were irradiated with 4 J/cm² on day 1, immediately after injury. Control lesions (CG) received no light and were left to heal spontaneously. Blood flow was measured on days 1, 3, 8, 10, 15, and 21 using laser Doppler flowmetry. Animals were killed on days 3, 8, 10, 15, and 21. Skin specimens were obtained and routinely processed for hematoxylin and eosin. The specimens were evaluated according to differential leukocyte counting and angiogenesis. Statistical analysis was performed, and significance was accepted at p < 0.05. Irradiated groups showed a peak of new vessels on day 15 while, for CG, the peak was on day 21. On day 21, FG exhibited a significantly greater number of cumulative neutrophils while SG showed a higher number of mononuclear cells. Our results confirm that both protocols used accelerate angiogenesis and stimulate leukocyte chemotaxis on burn treatment. In addition, this work suggests that a single-dose LLLT accelerates the inflammatory phase of skin repair.

Effect of laser therapy on skeletal muscle repair process in diabetic rats.

Skeletal muscle myopathy is a common source of disability in diabetic patients. This study evaluated whether low-level laser therapy (LLLT) influences the healing morphology of injured skeletal muscle. Sixty-five male Wistar rats were divided as follows: (1) sham; (2) control; (3) diabetic; (4) diabetic sham; (5) nondiabetic cryoinjured submitted to LLLT (LLLT); (6) diabetic cryoinjured submitted to LLLT (D-LLLT); and (7) diabetic cryoinjured non-treated (D). Diabetes was induced with streptozotocin. Anterior tibialis muscle was cryoinjured and received LLLT daily (780 nm, 5 J/cm(2), 10 s per point; 0.2 J; total treatment, 1.6 J). Euthanasia occurred on day 1 in groups 1, 2, 3, and 4 and on days 1, 7, and 14 in groups 5, 6, and 7. Muscle samples were processed for H&E and Picrosirius Red and photographed. Leukocytes, myonecrosis, fibrosis, and immature fibers were manually quantified using the ImageJ software. On day 1, all cryoinjured groups were in the inflammatory phase. The D group exhibited more myonecrosis than LLLT group (p < 0.05). On day 14, the LLLT group was in the remodeling phase; the D group was still in the proliferative phase, with fibrosis, chronic inflammation, and granulation tissue; and the D-LLLT group was in an intermediary state in relation to the two previous groups. Under polarized light, on day 14, the LLLT and D-LLLT groups had organized collagen bundles in the perimysium, whereas the diabetic groups exhibited fibrosis. LLLT can have a positive effect on the morphology of skeletal muscle during the tissue repair process by enhancing the reorganization of myofibers and the perimysium, reducing fibrosis.

[Atraumatic restorative treatment: a dental alternative well-received by children]. / La práctica restaurativa atraumática: una alternativa dental bien recibida por los niños.

The purpose of this study is to compare atraumatic restorative treatment (ART) with the conventional rotational restorative method (CM) to determine in both cases the total time required for the procedure, the cost, the presence of pain, and the behavior of pediatric patients in Peru. Of the 30 children selected for the study, half received ART and restoration with glass ionomer cement and the other half, CM and restoration with amalgam. The study parameters were the times required to remove the decayed tissue and to complete the entire procedure, the total cost of the procedure, the presence of pain, and the patient's behavior during treatment. Significant differences were found between the two techniques in all parameters, except for the patient's behavior. Although removing the decayed tissue was faster with the CM, the entire procedure was faster with ART, which, moreover, was significantly less expensive and less painful than the CM. The results indicated that ART is a very good alternative due to its low cost and acceptance by the children.

Vascular inflammation exposes perivascular cells to SARS-CoV-2 infection.

Pericytes stabilize blood vessels and promote vascular barrier function. However, vessels subjected to pro-inflammatory conditions have impaired barrier function, which has been suggested to potentially expose perivascular cells to SARS-CoV-2. To test this hypothesis, we engineered pericyte-supported vascular capillaries on-a-chip, and determined that the extravasation and binding of spike protein (S1) on perivascular cells of inflamed vessels to be significantly higher that in healthy controls, indicating a potential target to understand COVID-19 vascular complications.

In-vitro models of biocompatibility testing for restorative dental materials: From 2D cultures to organs on-a-chip.

Dental caries is a biofilm-mediated, diet-modulated, multifactorial and dynamic disease that affects more than 90% of adults in Western countries. The current treatment for decayed tissue is based on using materials to replace the lost enamel or dentin. More than 500 million dental restorations are placed annually worldwide, and materials used for these purposes either directly or indirectly interact with dentin and pulp tissues. The development and understanding of the effects of restorative dental materials are based on different in-vitro and in-vivo tests, which have been evolving with time. In this review, we first discuss the characteristics of the tooth and the dentin-pulp interface that are unique for materials testing. Subsequently, we discuss frequently used in-vitro tests to evaluate the biocompatibility of dental materials commonly used for restorative procedures. Finally, we present our perspective on the future directions for biological research on dental materials using tissue engineering and organs on-a-chip approaches. STATEMENT OF SIGNIFICANCE: Dental caries is still the most prevalent infectious disease globally, requiring more than 500 million restorations to be placed every year. Regrettably, the failure rates of such restorations are still high. Those rates are partially based on the fact that current platforms to test dental materials are somewhat inaccurate in reproducing critical components of the complex oral microenvironment. Thus, there is a collective effort to develop new materials while evolving the platforms to test them. In this context, the present review critically discusses in-vitro models used to evaluate the biocompatibility of restorative dental materials and brings a perspective on future directions for tissue-engineered and organs-on-a-chip platforms for testing new dental materials.

Histological and biochemical effects of preventive and therapeutic vascular photobiomodulation on rat muscle injury.

The intravascular or transcutaneous application of photobiomodulation (PBM) over blood vessels (vascular photobiomodulation, VPBM) has been used for the treatment of inflammatory and chronic conditions with promising systemic results. This study evaluated the VPBM effects on a model of muscle regeneration after acute injury and compared the outcomes of preventive and therapeutic VPBM. Transcutaneous VPBM was administered over the rat's main tail vein. Serum levels of creatine kinase (CK), aspartate aminotransferase (AST), and lactate were evaluated and muscles were processed for macroscopic and microscopic analysis. Preventive and therapeutic VPBM led to decreased inflammatory infiltrate, edema, and myonecrosis but with an increase in immature muscle fibers. CK, AST, and lactate levels were lower in the groups treated with VPBM (lowest concentrations in preventive VPBM application). Preventive and therapeutic VPBM were capable of exerting a positive effect on acute muscle injury repair, with more accentuated results when preventive VPBM was administered.

A dual-ink 3D printing strategy to engineer pre-vascularized bone scaffolds in-vitro.

A functional vascular supply is a key component of any large-scale tissue, providing support for the metabolic needs of tissue-remodeling cells. Although well-studied strategies exist to fabricate biomimetic scaffolds for bone regeneration, success rates for regeneration in larger defects can be improved by engineering microvascular capillaries within the scaffolds to enhance oxygen and nutrient supply to the core of the engineered tissue as it grows. Even though the role of calcium and phosphate has been well understood to enhance osteogenesis, it remains unclear whether calcium and phosphate may have a detrimental effect on the vasculogenic and angiogenic potential of endothelial cells cultured on 3D printed bone scaffolds. In this study, we presented a novel dual-ink bioprinting method to create vasculature interwoven inside CaP bone constructs. In this method, strands of a CaP ink and a sacrificial template material was used to form scaffolds containing CaP fibers and microchannels seeded with vascular endothelial and mesenchymal stem cells (MSCs) within a photo-crosslinkable gelatin methacryloyl (GelMA) hydrogel material. Our results show similar morphology of growing vessels in the presence of CaP bioink, and no significant difference in endothelial cell sprouting was found. Furthermore, our initial results showed the differentiation of hMSCs into pericytes in the presence of CaP ink. These results indicate the feasibility of creating vascularized bone scaffolds, which can be used for enhancing vascular formation in the core of bone scaffolds.

Engineering pericyte-supported microvascular capillaries in cell-laden hydrogels using stem cells from the bone marrow, dental pulp and dental apical papilla.

Engineered tissue constructs require the fabrication of highly perfusable and mature vascular networks for effective repair and regeneration. In tissue engineering, stem cells are widely employed to create mature vascularized tissues in vitro. Pericytes are key to the maturity of these vascular networks, and therefore the ability of stem cells to differentiate into pericyte-like lineages should be understood. To date, there is limited information regarding the ability of stem cells from the different tissue sources to differentiate into pericytes and form microvascular capillaries in vitro. Therefore, here we tested the ability of the stem cells derived from bone marrow (BMSC), dental pulp (DPSC) and dental apical papilla (SCAP) to engineer pericyte-supported vascular capillaries when encapsulated along with human umbilical vein endothelial cells (HUVECs) in gelatin methacrylate (GelMA) hydrogel. Our results show that the pericyte differentiation capacity of BMSC was greater with high expression of α-SMA and NG2 positive cells. DPSC had α-SMA positive cells but showed very few NG2 positive cells. Further, SCAP cells were positive for α-SMA while they completely lacked NG2 positive cells. We found the pericyte differentiation ability of these stem cells to be different, and this significantly affected the vasculogenic ability and quality of the vessel networks. In summary, we conclude that, among stem cells from different craniofacial regions, BMSCs appear more suitable for engineering of mature vascularized networks than DPSCs or SCAPs.