Figure 4 - uploaded by Takashi I
Content may be subject to copyright.
Tissue regenerative activity and detection of donor cells at day 7 postirradiation. The photomicrograph on the upper left corner shows a hematoxylin and eosin staining of a tongue. The red box represents the area of the ulcer and is shown in higher magnification; the blue box represents the area of the mucositis adjacent to the ulcer and is shown in higher magnification (×15). (A) Cell proliferation (PCNA) immunostaining of ulcerated area at day 7 postirradiation. Three blue boxes in the upper panels (×100) are shown in higher magnification in the lower panels (scale bar is 50 µm) (×200). PCNA-positive cells (stained in brown) were mainly detected in the basal layer of the epithelium. (b) Number of PCNA-positive cells in epithelium (mean value ± SD). (c) Blood vessel area in dermis just below the basement membrane (mean value ± SD). (D) vWF immunofluorescence staining of the mucositis area adjacent to ulcer at day 7. Three white boxes in the upper panels (×100) are shows in higher magnification in the lower panels (scale bar is 50 µm) (×200). Abundant blood vessel formation (stained in red) is shown under the basement membrane of the BM group. (E) EGFR immunofluorescence staining (red) of the lingual epithelium in mucositis area adjacent to ulcer area on 2, 5, and 7 d postirradiation (scale bar is 25 µm) (×400). (F) Detection of donor BMDCs in tongues at day 7 postirradiation. Expression of sry mRNA (on Y chromosome) in female mice (reverse transcription polymerase chain reaction) and detection of PKH26-labeled cells (red) were assessed. Sry gene expression was not detected in the female mice of the IR group. Sry gene was detected in female mice transplanted with male BMDCs. PKH26-labeled cells were detected right under the basement membrane adjacent to the ulcer area in the BM group (scale bar is 50 µm) (×200). IV, intravenous injection; BM group; blue, DAPI; yellow-dotted line, boundary of the epithelium and dermis. Red: PKH26-labeled cells. Black asterisk represents statistical significance between control and both irradiated groups (p < .05). Red asterisk represents statistical significance between IR and BM groups (p < .05) (mean ± SD). BMDC, bone marrow-derived cell; Contl, control group; IR, irradiation + no cell transplant group; BM, irradiation + bone marrow-derived cells transplantation group.
Source publication
Oral mucositis (ulcer) is a serious and painful side effect for patients with head and neck cancer following radiation therapy. However, current clinical strategies cannot efficiently prevent the occurrence of oral muco-sitis. In this study, we investigated whether bone marrow-derived cells (BMDCs) prevented the occurrence and/or decreased the seve...
Contexts in source publication
Context 1
... the maximum ulceration period (day 7 postirradiation), mucositis area (including ulcer) in tongues was recognized by toluidine blue staining (Figure 1B and Appendix Figures 3, 4A). When the mucositis area was quantified, its surface area, including ulcers in tongues of the BM group (9.4%), was significantly lower than that of the IR group (56.7%) and the dermal fibroblast group (62.5%; Figure 1B, 1C and Appendix Figure 4B). Furthermore, the area of only ulcers in tongues of the BM group (6.8%) was also markedly lower when compared with that of the IR group (23.1%) and the dermal fibroblast group (18.3%; Figure 1B, 1D and Appendix Figure 4C). ...
Context 2
... the mucositis area was quantified, its surface area, including ulcers in tongues of the BM group (9.4%), was significantly lower than that of the IR group (56.7%) and the dermal fibroblast group (62.5%; Figure 1B, 1C and Appendix Figure 4B). Furthermore, the area of only ulcers in tongues of the BM group (6.8%) was also markedly lower when compared with that of the IR group (23.1%) and the dermal fibroblast group (18.3%; Figure 1B, 1D and Appendix Figure 4C). ...
Context 3
... cells were abundantly detected in the epithelial basal layer of nonirradiated control mice. There was a smaller number of PCNA-positive cells at the ulcer area of the IR and BM groups at 7 d postirradiation ( Figure 4A). However, basal cells in the BM group had high PCNA activity, although there was no significant difference compared with those cells in the IR group (p = .105; ...
Context 4
... basal cells in the BM group had high PCNA activity, although there was no significant difference compared with those cells in the IR group (p = .105; Figure 4A, 4B). Moreover, blood vessel formation was more abundant in the dermis below the basement membrane adjacent to the ulcer area of the BM group when compared with that of the control and IR groups ( Figure 4C, 4D). ...
Context 5
... 4A, 4B). Moreover, blood vessel formation was more abundant in the dermis below the basement membrane adjacent to the ulcer area of the BM group when compared with that of the control and IR groups ( Figure 4C, 4D). ...
Context 6
... further evaluate tissue regenerative activity, expression of EGFR (which has crucial roles for epithelium restoration) in the epithelium adjacent to ulcer area was assessed. In the BM group, EGFR expression was observed from 2 to 5 d postirradiation ( Figure 4E and Appendix Figure 5E). However, this expression almost disappeared at 7 d. ...
Context 7
... day 7 postirradiation, Y chromosome signal (Sry gene derived from donor male cells) was detected in harvested tongues of the BM group as well as in control male mice ( Figure 4F). However, this signal was not found in female mice of the IR group. ...
Citations
... What's more, during radiotherapy, inflammation, including radiation-induced mucositis, may occur [49][50][51] and increases energy expenditure and breaks down protein [52], with the loss of muscle mass [53]. According to Unsal et al. after radiotherapy, the malnutrition rate of patients with head and neck cancer is 88% [54]. ...
Background
Malnutrition is a common complication in patients with nasopharyngeal carcinoma (NPC). However, there are few studies on risk factors for malnutrition in NPC patients. Our aims were to identify the risk factors for malnutrition in NPC patients.
Methods
NPC patients were recruited in this cross-sectional study, and they were divided into well-nourished and malnourished groups according to the Global Leadership Initiative on Malnutrition (GLIM). Potential risk factors were initially screened using univariate analysis (p < 0.1), and the selected ones were analyzed by logistic regression analysis (p < 0.05) to identify the risk factors for malnutrition in NPC patients.
Results
In total, 305 NPC patients meeting eligibility criteria were enrolled. Multivariate logistic regression analysis revealed that low body mass index (BMI) (OR = 0.596, 95% CI 0.520–0.683, p < 0.001), the high total radiation dose received (OR = 1.046, 95% CI 1.023–1.069, p < 0.001), appetite loss (OR = 2.839, 95% CI 1.269–6.353, p = 0.011), and low PA (OR = 0.993, 95% CI 0.988–0.998, p = 0.008) were risk factors for malnutrition in NPC patients.
Conclusions
The low BMI, the high total radiation dose received, appetite loss, and low prealbumin were risk factors for malnutrition in NPC patients.
... Improvement in bone marrow transplantation not only restores hematopoietic function but also alleviates other digestive symptoms (129,130). Bone marrow-derived cells can also reduce radiogenic oral mucositis (131). To further determine how bone marrow restores digestive symptoms, Tran et al. injected bone marrow soluble extract ("soup") into mice and found that bone marrow soup restored salivary flow rates to normal levels; protected salivary acinar, ductal, myoepithelial, and progenitor cells; increased cell proliferation and blood vessels; and upregulated the expression of tissue remodeling/repair/ regenerative genes. ...
Radiotherapy is one of the main therapeutic methods for treating cancer. The digestive system consists of the gastrointestinal tract and the accessory organs of digestion (the tongue, salivary glands, pancreas, liver and gallbladder). The digestive system is easily impaired during radiotherapy, especially in thoracic and abdominal radiotherapy. In this review, we introduce the physical classification, basic pathogenesis, clinical characteristics, predictive/diagnostic factors, and possible treatment targets of radiotherapy-induced digestive injury. Radiotherapy-induced digestive injury complies with the dose-volume effect and has a radiation-based organ correlation. Computed tomography (CT), MRI (magnetic resonance imaging), ultrasound (US) and endoscopy can help diagnose and evaluate the radiation-induced lesion level. The latest treatment approaches include improvement in radiotherapy (such as shielding, hydrogel spacers and dose distribution), stem cell transplantation and drug administration. Gut microbiota modulation may become a novel approach to relieving radiogenic gastrointestinal syndrome. Finally, we summarized the possible mechanisms involved in treatment, but they remain varied. Radionuclide-labeled targeting molecules (RLTMs) are promising for more precise radiotherapy. These advances contribute to our understanding of the assessment and treatment of radiation-induced digestive injury.
... Mice were sacrificed and the whole tongue was then removed from oral cavity. Excised tongues were stained with 0.05% toluidine blue to visualize the ulceration (19). Excised tongues of mice were stained with 0.05% toluidine blue for 10 min and rinsed with 10% acetic acid for 1 min to visualize the ulceration (19). ...
... Excised tongues were stained with 0.05% toluidine blue to visualize the ulceration (19). Excised tongues of mice were stained with 0.05% toluidine blue for 10 min and rinsed with 10% acetic acid for 1 min to visualize the ulceration (19). Ulcers were visible as a deep blue color after staining. ...
Background
Oral mucositis is the most common oral complication of cancer patients receiving radiotherapy and/or chemotherapy, leading to poor quality of life. Limitations of the current interventions on radiation-induced oral mucositis (RIOM) urge the development of novel therapeutics. Here, we evaluated the treatment outcome of probiotic Streptococcus salivarius K12 on RIOM mice, and oral microbiota that is associated with the progress of RIOM was further investigated.
Methods
An experimental RIOM mouse model was established, and S. salivarius K12 was applied to the mouse oral cavity daily. Histological analyses were performed to evaluate the severity of oral mucositis and the treatment outcome of S. salivarius K12. The oral microbiota of mice was further analyzed by 16S rRNA sequencing, microbial culture and qPCR.
Results
Irradiation induced conspicuous mucositis in the oral cavity of mice. S. salivarius K12 treatment was beneficial for the healing of RIOM, as reflected by reduced ulcer size, increased basal layer epithelial cellularity and mucosal thickness, and elevated epithelial proliferation and attenuated apoptosis. RIOM mice presented significant oral microbial dysbiosis, with an overgrowth of oral anaerobes. S. salivarius K12 treatment reconstituted the oral microbiota and decreased the abundance of oral anaerobes of RIOM mice. In addition, S. salivarius K12 treatment inhibited NI1060 in Pasteurella genus and downregulated the expression of nitrate reductase.
Conclusions
S. salivarius K12 treatment can alleviate RIOM and reconstituted the dysbiotic oral microbiota in mice. S. salivarius K12 may represent a promising adjuvant treatment to improve the quality of life of cancer patients receiving radiotherapy.
... 48 Only eight original studies primarily concerned tongue regeneration. [49][50][51][52][53][54][55][56] This is surprising given fibrosis or loss of tongue volume commonly follows radiotherapy and surgery for upper aerodigestive tract cancer and significantly affects phonation and swallow. Cell therapy procedures and small molecules that can overcome fibrosis, as with the vocal folds, would benefit patients and would be relatively easy to deploy endoscopically as a graft or injectable. ...
Objective
This review assesses regenerative medicine of the upper aerodigestive tract during the first two decades of the twenty-first century, focusing on end-stage fibrosis and tissue loss in the upper airways, salivary system, oropharynx and tongue.
Method
PubMed, Embase, Google Scholar, Cochrane Library, Medline and clinicaltrials.org were searched from 2000 to 2019. The keywords used were: bioengineering, regenerative medicine, tissue engineering, cell therapy, regenerative surgery, upper aerodigestive tract, pharynx, oropharynx, larynx, trachea, vocal cord, tongue and salivary glands. Original studies were subcategorised by anatomical region. Original human reports were further analysed. Articles on periodontology, ear, nose and maxillofacial disorders, and cancer immunotherapy were excluded.
Results
Of 716 relevant publications, 471 were original studies. There were 18 human studies included, within which 8 reported airway replacements, 5 concerned vocal fold regeneration and 3 concerned salivary gland regeneration. Techniques included cell transplantation, injection of biofactors, bioscaffolding and bioengineered laryngeal structures.
Conclusion
Moderate experimental success was identified in the restoration of upper airway, vocal fold and salivary gland function. This review suggests that a shift in regenerative medicine research focus is required toward pathology with a higher disease burden.
... Radiation stomatitis is a radiationinduced disease that generally occurs in patients who have received radiotherapy for head or neck cancer. Severe radiation stomatitis can be seen in 29 to 66% of all head and neck cancer patients after undergoing radiotherapy (Sumita et al., 2014). The extremely painful symptoms associated with these ulcerative diseases may lead to difficulty in chewing, swallowing, and speaking. ...
Patients in dental hospitals often experience oral ulcerative lesions, which lead to pain and affect the patient's quality of life. At present, the goal of treating oral ulcerative lesions with drugs is to reduce inflammation and promote ulcer healing. However, very few antibacterial and hemostatic drugs are designed to be suitable for the microenvironment of gingival ulcers. Based on this, we have designed a natural therapeutic agent for oral ulcerative lesions that meets the various requirements of oral ulcerative lesion medication. The chitosan-g-polyacrylamide (CP) copolymer is composed of chitosan as the main chain and polyacrylamide polymers as the side chains. Antibacterial experiments show that this polymer can effectively inhibit the proliferation of Gram-negative (Escherichia coli) and Gram-positive bacteria (Staphylococcus aureus). In vitro cell experiments also show that the CP copolymer is non-toxic, which is conducive to ulcer wound healing. Coagulation experiments prove that the CP copolymer can accelerate blood coagulation to stop bleeding. In experiments using a Wistar rat gingival ulcer model, the CP copolymer significantly promoted ulcer healing and shortened the healing time. These results indicate that the CP copolymer may serve as a potential therapeutic agent for oral ulcerative lesions.
... This study is a pre-requisite step for future clinical trials aimed at developing cell-based therapies for atrophic SGs. Although the mechanisms of regeneration are not currently well understood, we have previously found that autologous BMDC injections have beneficial therapeutic effects via the promotion of anti-inflammation and vasculogenesis in radiogenic-injured SGs or oral mucositis mouse models [6,27]. Therefore, the use of E-MNCs, which do not require a long cell expansion process, is a simple and direct approach using a readily available source of cells (blood) that can be obtained with low invasiveness. ...
Background:
There are currently no effective treatments available for patients with irreversible loss of salivary gland (SG) function caused by radiation therapy for head and neck cancer. In this study, we have developed an effective culture method to enhance the anti-inflammatory and vasculogenic phenotypes of peripheral blood mononuclear cells (PBMNCs) and investigated whether such effectively conditioned PBMNCs (E-MNCs) could regenerate radiation-injured SGs and ameliorate salivary secretory function in mice.
Methods:
Mouse PBMNCs were expanded in primary serum-free culture with five vasculogenic proteins for 5 days, and then the resulting cells (E-MNCs) were analyzed for their characteristics. Subsequently, 5 × 104 E-MNCs (labeled with EGFP in some experiments) were injected intra-glandularly into a mouse model of radiation-injured atrophic submandibular glands. After 2-3 weeks, the submandibular glands were harvested, and then the injected E-MNCs were tracked. Four, 8, and 12 weeks after irradiation (IR), salivary outputs were measured to evaluate the recovery of secretory function, and the gland tissues were harvested for histological and gene expression analyses to clarify the effects of cell transplantation.
Results:
The resulting E-MNCs contained an enriched population of definitive CD11b/CD206-positive (M2 macrophage-like) cells and showed anti-inflammatory and vasculogenic characteristics. Salivary secretory function in E-MNC-transplanted mice gradually recovered after 4 weeks post-irradiation (post-IR) and reached 3.8-fold higher than that of non-transplanted mice at 12 weeks. EGFP-expressing E-MNCs were detected in a portion of the vascular endothelium and perivascular gland tissues at 2 weeks post-IR, but mainly in some microvessels at 3 weeks. Between 4 and 12 weeks post-IR, mRNA expression and histological analyses revealed that E-MNC transplantation reduced the expression of inflammatory genes and increased the level of tissue-regenerative activities such as stem cell markers, cell proliferation, and blood vessel formation. At 12 weeks post-IR, the areas of acinar and ductal cells regenerated, and the glands had less fibrosis.
Conclusions:
This effective conditioning of PBMNCs is a simple, rapid, and efficient method that provides a non-invasive source of therapeutic cells for regenerating radiation-injured atrophic SGs.
... These findings are consistent with the steady increase of epithelial cells proliferation noticed from the beginning of the experimental period till its end. This is may be explained by the following first; the paracrine effect of the transplanted stem cells through growth factors and cytokines, which they release to increase epithelial cells proliferation (31,32) and to make the microenvironment more appropriate for the repair of injury (33). Second; by transdifferentiating into keratinocytes directly (34,35). ...
Background and objectives:
Oral ulceration is one of the most common debilitating condition that affects the oral cavity. In this study, the effect of locally injected platelet rich plasma (PRP) and bone marrow-derived mesenchymal stem cells (BMSCs) on the healing of oral ulcer was investigated.
Methods and results:
An ulcer was induced in buccal mucosa of rats by using 5mm biopsy punch followed by application of cotton swab soaked with formocresol for 60sec. The ulcer was left untreated in the control group, treated with intralesional injection of PRP, or isolated cultured BMSCs. Data were analyzed clinically, histologically and immunohistologically on day 3, 5, 7 and 10. BMSCs group showed smaller ulcer area throughout the whole experimental period than the other groups with complete resolution of the ulcer on day 10, unlike the control group. However, there was no significant difference with PRP, on day 5, 7 and 10, regarding clinical ulcer size. BMSCs group showed better histological results regarding the rate of epithelial cell migration, the number of inflammatory cells, thickness and organization of collagen fibres and the number of blood vessels, with complete re-epithelization on day 10. BMSCs group showed a greater number of anti-PCNA positive nuclei throughout the whole experimental period than the other groups except on day 5, PRP had higher mean numbers of anti-PCNA positive nuclei in both tissues.
Conclusions:
Both PRP and BMSCs accelerate wound healing and enhance the quality of the healing tissue with the latter being slightly more effective and faster.
... All the experiments were approved by the Animal Care and Use Committee of Sun Yat-sen University (IACUC-DB-16-1215). Animal models were established as previously described 42 . The tongue was extended out of the mouth and exposed to 16 Gy (a dose of 1.6 Gy/min) of radiation through a 10-mm-diameter hole to establish RIM. ...
Radiation-induced oral mucositis affects patient quality of life and reduces tolerance to cancer therapy. Unfortunately, traditional treatments are insufficient for the treatment of mucositis and might elicit severe side effects. Due to their immunomodulatory and anti-inflammatory properties, the transplantation of mesenchymal stem cells (MSCs) is a potential therapeutic strategy for mucositis. However, systemically infused MSCs rarely reach inflamed sites, impacting their clinical efficacy. Previous studies have demonstrated that chemokine axes play an important role in MSC targeting. By systematically evaluating the expression patterns of chemokines in radiation/chemical-induced oral mucositis, we found that CXCL2 was highly expressed, whereas cultured MSCs negligibly express the CXCL2 receptor CXCR2. Thus, we explored the potential therapeutic benefits of the transplantation of CXCR2-overexpressing MSCs (MSCsCXCR2) for mucositis treatment. Indeed, MSCsCXCR2 exhibited enhanced targeting ability to the inflamed mucosa in radiation/chemical-induced oral mucositis mouse models. Furthermore, we found that MSCCXCR2 transplantation accelerated ulcer healing by suppressing the production of pro-inflammatory chemokines and radiogenic reactive oxygen species (ROS). Altogether, these findings indicate that CXCR2 overexpression in MSCs accelerates ulcer healing, providing new insights into cell-based therapy for radiation/chemical-induced oral mucositis.
... Pain caused by oral mucositis may hinder proper nutrition, which in turn leads to weight loss, cachexia, and dehydration of the body, which is already weakened by disease. [1][2][3] Pain caused by mucositis also affects drug administration, speech, and breathing. [4][5][6] Oral mucosa may become deteriorated to such an extent that the patients require changes in the antitumor treatment and/or administration of parenteral analgesia. ...
... 25,26 In an effort to minimize the side effects of radiation therapy and chemotherapy used in cancer treatment, there have been studies aimed at developing therapeutic strategies to regenerate oral mucosal tissues affected by mucositis. 3,5 Certain agents have been examined for prevention or treatment of oral mucositis caused by chemotherapy or radiotherapy, but none of them has been confirmed as fully effective. 27 A study by Pereira Pinto et al. revealed that in a group of children with ALL who used 0.12% chlorhexidine gluconate to rinse the mouth, 26% developed mucositis in comparison with 80% who did not use the oral rinse. ...
Background:
Oral mucositis is a problem occurring within the oral cavity, which is the most difficult to deal with during anti-tumor treatment. The first symptom reported by the patient is discomfort. Salivary immunoglobulins play an important role in pathological processes occurring in the oral cavity.
Objectives:
The study objective was to assess the occurrence of oral mucositis and to assess changes in the saliva IgA, IgG, and IgM concentration in children with acute lymphoblastic leukemia during antitumor treatment.
Material and methods:
The study included 78 children with acute lymphoblastic leukemia (ALL) and a control group of healthy children. All the participants underwent 3 examinations.
Results:
Mucosal opacity followed by redness usually occurred within 2-4 days after the methotrexate infusion. The most severe lesions of the oral mucosa were observed after the 1st month of chemotherapy. Correlations were found between hard-to-heal wounds and ulcers and blood morphology parameters. The mean saliva IgA concentration in children with ALL during chemotherapy was significantly lower than in children in the control group. A comparison of the mean saliva IgG in a given patient in subsequent examinations revealed a significant saliva IgG decrease occurring between the 1st and 3rd examinations.
Conclusions:
Wounds and ulcers that were difficult to heal were related to blood morphology parameters. A low salivary IgA concentration in children with ALL may result in the development and potentiation of oral lesions typical of mucositis during anti-tumor treatment. A significant decrease in salivary IgG and IgM concentrations in children with ALL during chemotherapy may cause potentiation of pathological lesions in the oral mucosa.
... On the other hand, new possibilities for minimally invasive and non-laborious procedures using fresh non-cultured cells obtained from small amounts of autologous tissues, such as bone marrow, adipose, and dermal tissues, have been suggested from recent research on cell-based therapies in tissue engineering (I et al., 2014;Sumita et al., 2011;Yoo and Lim, 2009). For example, it was recently reported that the transplantation of micronized tissues (micro-graft) 50 µm in size from small autologous tissue samples (only a few millimeters in size) can be used effectively for bone or soft tissue engineering (Giaccone, Brunetti, Camandona, Trovato, & Graziano, 2014;Marcarelli, Trovato, Novarese, Riccio, & Graziano, 2017;Monti et al., 2017;Svolacchia, De Francesco, Trovato, Graziano, & Ferraro, 2016;Trovato et al., 2015;Zanzottera, Lavezzari, Trovato, Icardi, & Graziano, 2014). ...
The free gingival graft (FGG) and connective tissue graft (CTG) are currently considered to be the gold standards for keratinized gingival tissue reconstruction and augmentation. However, these procedures have some disadvantages in harvesting large grafts, such as donor-site morbidity as well as insufficient gingival width and thickness at the recipient site post-treatment. To solve these problems, we focused on an alternative strategy using micronized tissue transplantation (micro-graft). In this study, we first investigated whether transplantation of micronized gingival connective tissues (MGCTs) promotes skin wound healing. MGCTs (≤100 µm) were obtained by mincing a small piece (8 mm(3) ) of porcine keratinized gingiva using the RIGENERA system. The MGCTs were then transplanted to a full skin defect (5 mm in diameter) on the dorsal surface of immunodeficient mice after seeding to an atelocollagen matrix. Transplantations of atelocollagen matrixes with and without micronized dermis were employed as experimental controls. The results indicated that MGCTs markedly promote the vascularization and epithelialization of the defect area 14 days after transplantation compared to the experimental controls. After 21 days, complete wound closure with low contraction was obtained only in the MGCT grafts. Tracking analysis of transplanted MGCTs revealed that some mesenchymal cells derived from MGCTs can survive during healing and may function to assist in wound healing. We propose here that micro-grafting with MGCTs represents an alternative strategy for keratinized tissue reconstruction that is characterized by low morbidity and ready availability. This article is protected by copyright. All rights reserved.
