<strong>Purpose: </strong>To establish a simple and accurate photodynamic diagnosis (PDD) method for oral squamous cell carcinoma (OSCC). <strong>Methods: </strong>OSCC cell lines HSC-2, HSC-3,...<strong>Purpose: </strong>To establish a simple and accurate photodynamic diagnosis (PDD) method for oral squamous cell carcinoma (OSCC). <strong>Methods: </strong>OSCC cell lines HSC-2, HSC-3, HSC-4, and Sa3, and normal human oral keratinocytes (HOK) were used. First, we examined the amount of cells needed to detect differences in fluorescence intensities for PDD. OSCC cell lines were adjusted to concentrations of 1 × 10<sup>4</sup> (10<sup>4</sup>), 1 × 10<sup>5</sup> (10<sup>5</sup>), and 1 × 10<sup>6</sup> (10<sup>6</sup>) cells/ml. The experimental groups comprised a group with 5-aminolevulinic acid (5-ALA (+)), and a group without 5-ALA (5-ALA (-)). For each OSCC cell line, 100 μl of each concentration of cells of the 5-ALA groups was seeded onto fluorescence plates, and fluorescence intensity was measured at 60-min intervals for 240 min. Results are expressed as the ratio of fluorescence intensity in 5-ALA (+) to 5-ALA (-). As cells at the concentration of 10<sup>6</sup> cells/ml provided the clearest results, fluorescence intensities of all cell lines were measured using this concentration at 20-min intervals for 700 min using the same methods. <strong>Results: </strong>The 5-ALA (+) to (-) ratio increased in a cell concentration-dependent manner at 240 min;the ratio was highest with 10<sup>6</sup> cells/ml and lowest with 10<sup>4</sup> cells/ml. With 10<sup>6</sup> cells/ml in the 5-ALA (+) group, fluorescence intensity increased in a metabolic time-dependent manner;the increase was highest in HSC-2 cells, followed by HSC-4 cells, HSC-3 cells, Sa3 cells, and HOK. Fluorescence intensity was significantly enhanced after 40 min in HSC-2, HSC-3, and HSC-4 cells, after 60 min in Sa3 cells, and after 100 min in HOK compared to the 5-ALA (-) group (<em>P </em>< 0.05). Moreover, fluorescence intensity was significantly increased in OSCC cell lines compared to HOK after 40 min. <strong>Conclusion:</strong> Early detection of OSCC is possible by screening only microplate reader measurements of fluorescence intensity for PDD.展开更多
A series of 5-aminolevulinic acid and its alkylester methanesulfonates was exploited to photodynamic therapy（PDT） of human lymphocytic cells, U-937 in vitro. The PDT efficiency is influenced by the concentration and...A series of 5-aminolevulinic acid and its alkylester methanesulfonates was exploited to photodynamic therapy（PDT） of human lymphocytic cells, U-937 in vitro. The PDT efficiency is influenced by the concentration and incubation time. Generally, for ALA and its alkylester methanesulfonates, the cell survival rate decreases and the accumulation ability of PplX increases with the concentration and incubation time. We found that the longer carbon chain methanesulfonates（C5-S, C6-S, C8-S） exhibit better PDT effect than ALA methanesulfonate. This possibly provides a promising route to the clinical application ofPplX-mediated PDT to cancer cell.展开更多
Objective To optimize experimental parameters for the photosensitization of 5-aminolevulinic acid (ALA) in promyelocytic leukemia cell HL60 and compare them with normal human peripheral blood mononuclear cell (PBMC). ...Objective To optimize experimental parameters for the photosensitization of 5-aminolevulinic acid (ALA) in promyelocytic leukemia cell HL60 and compare them with normal human peripheral blood mononuclear cell (PBMC). Methods ALA incubation time, wavelength applied to irradiate, concentration of ALA incubated, irradiation fluence may modulate the effect of 5-aminolevulinic acid based Photodynamic Therapy (ALA-PDT).The high-pressure mercury lamps of 400W served as light source, the interference filter of 410nm, 432nm, 545nm, 577nm were used to select the specific wavelength. Fluorescence microscope was used to detect the fluorescence intensity and location of protoporphyrin IX (PpIX) endogenously produced by ALA. MTT assay was used to measure the survival of cell. Flow cytometry with ANNEXIN V FITC kit (contains annexin V FITC, binding buffer and PI) was used to detect the mode of cell death. Results ① 1mmol/L ALA incubated 1×105/mL HL60 cell line for 4 hours, the maximum fluorescence of ALA induced PpIX was detected in cytomembrane. ② Irradiated with 410nm for 14.4J/cm2 can result in the minimum survivability of HL60 cell. ③ The main mode of HL60 cell death caused by ALA-PDT is necrosis. Conclusion ALA for 1mmol/L, 4 hours for dark incubation time, 410nm for irradiation wavelength, 14.4J/cm2 for irradiation fluence were the optimal parameters to selectively eliminate promyelocytic leukemia cell HL60 by ALA based PDT. The photosensitization of ALA based PDT caused the necrosis of HL60 cell, so it could be used for inactivation of certain leukemia cells.展开更多
Objective:The aim of the study was to compare the effects of photodynamic therapy(PDT) with δ-aminolevulinic acid(ALA) for patients with different kinds of skin cancers and pre-cancers.Methods:The present study enrol...Objective:The aim of the study was to compare the effects of photodynamic therapy(PDT) with δ-aminolevulinic acid(ALA) for patients with different kinds of skin cancers and pre-cancers.Methods:The present study enrolled seventyfive cases,which included 17 cases of actinic keratosis(AK),9 cases of Bowen's disease,11 cases of superficial basal cell carcinomas(BCC),23 cases of nodule basal cell carcinomas and 15 cases of squamous cell carcinomas(SCC),and every patient had single lesion.All patients were treated with 20% ALA topically and He-Ne laser weekly for three times,and followed up 1-3 years.Results:After therapy,the rates of complete reaction(CR) were 100% in AK lesions,77.8% in Bowen's diseases,90.9% in superficial BCCs,47.8% in nodule BCCs,and 50.3% in SCCs,which had significant differences among these five kinds of lesions(H = 18.27,P < 0.05).The therapeutic effectiveness of ALA-PDT for AK was superior to that of Bowen's disease(Q = 4.364,P < 0.05),superficial BCC(Q = 5.55,P < 0.01),SCC(Q = 8.94,P < 0.01) and nodule BCC(Q = 17.91,P < 0.01);the effect of Bowen's disease was better than that of SCC(Q = 7.8,P < 0.01),nodule BCC(Q = 13.44,P < 0.01);the effect of superficial BCC was better than that of SCC(Q = 9.73,P < 0.01),nodule BCC(Q = 16.28,P < 0.01),but similar with Bowen's disease(Q = 0.96,P > 0.05);the effect of SCC was better than that of nodule BCC(Q = 17.74,P < 0.01).Conclusion:Our study shows that therapeutic effectiveness of ALA-PDT for AK is best in five diseases,and Bowen's disease and superficial BCC are secondary,while nodule BCC and SCC are at the bottom.展开更多
Background:Epidermal stem cells(EpSCs)that reside in cutaneous hair follicles and the basal layer of the epidermis are indispensable for wound healing and skin homeostasis.Little is known about the effects of photoche...Background:Epidermal stem cells(EpSCs)that reside in cutaneous hair follicles and the basal layer of the epidermis are indispensable for wound healing and skin homeostasis.Little is known about the effects of photochemical activation on EpSC differentiation,proliferation and migration during wound healing.The present study aimed to determine the effects of photodynamic therapy(PDT)on wound healing in vivo and in vitro.Methods:We created mouse full-thickness skin resection models and applied 5-aminolevulinic acid(ALA)for PDT to the wound beds.Wound healing was analysed by gross evaluation and haematoxylin–eosin staining in vivo.In cultured EpSCs,protein expression was measured using flow cytometry and immunohistochemistry.Cell migration was examined using a scratch model;apoptosis and differentiation were measured using flow cytometry.Results:PDT accelerated wound closure by enhancing EpSC differentiation,proliferation and migration,thereby promoting re-epithelialization and angiogenesis.PDT inhibited inflammatory infiltration and expression of proinflammatory cytokines,whereas the secretion of growth factors was greater than in other groups.The proportion of transient amplifying cells was significantly greater in vivo and in vitro in the PDT groups.EpSC migration was markedly enhanced after ALAinduced PDT.Conclusions:Topical ALA-induced PDT stimulates wound healing by enhancing re-epithelialization,promoting angiogenesis as well as modulating skin homeostasis.This work provides a preliminary theoretical foundation for the clinical administration of topical ALA-induced PDT in skin wound healing.展开更多
文摘<strong>Purpose: </strong>To establish a simple and accurate photodynamic diagnosis (PDD) method for oral squamous cell carcinoma (OSCC). <strong>Methods: </strong>OSCC cell lines HSC-2, HSC-3, HSC-4, and Sa3, and normal human oral keratinocytes (HOK) were used. First, we examined the amount of cells needed to detect differences in fluorescence intensities for PDD. OSCC cell lines were adjusted to concentrations of 1 × 10<sup>4</sup> (10<sup>4</sup>), 1 × 10<sup>5</sup> (10<sup>5</sup>), and 1 × 10<sup>6</sup> (10<sup>6</sup>) cells/ml. The experimental groups comprised a group with 5-aminolevulinic acid (5-ALA (+)), and a group without 5-ALA (5-ALA (-)). For each OSCC cell line, 100 μl of each concentration of cells of the 5-ALA groups was seeded onto fluorescence plates, and fluorescence intensity was measured at 60-min intervals for 240 min. Results are expressed as the ratio of fluorescence intensity in 5-ALA (+) to 5-ALA (-). As cells at the concentration of 10<sup>6</sup> cells/ml provided the clearest results, fluorescence intensities of all cell lines were measured using this concentration at 20-min intervals for 700 min using the same methods. <strong>Results: </strong>The 5-ALA (+) to (-) ratio increased in a cell concentration-dependent manner at 240 min;the ratio was highest with 10<sup>6</sup> cells/ml and lowest with 10<sup>4</sup> cells/ml. With 10<sup>6</sup> cells/ml in the 5-ALA (+) group, fluorescence intensity increased in a metabolic time-dependent manner;the increase was highest in HSC-2 cells, followed by HSC-4 cells, HSC-3 cells, Sa3 cells, and HOK. Fluorescence intensity was significantly enhanced after 40 min in HSC-2, HSC-3, and HSC-4 cells, after 60 min in Sa3 cells, and after 100 min in HOK compared to the 5-ALA (-) group (<em>P </em>< 0.05). Moreover, fluorescence intensity was significantly increased in OSCC cell lines compared to HOK after 40 min. <strong>Conclusion:</strong> Early detection of OSCC is possible by screening only microplate reader measurements of fluorescence intensity for PDD.
基金Supported Partly by the Academic Frontier Promotion Grant from the Ministry of Education,Science,Sports and Culture,Japan and the Exchange Program Between the Japan Society of the Promotion of Science and the Chinese Academy of Sciences
文摘A series of 5-aminolevulinic acid and its alkylester methanesulfonates was exploited to photodynamic therapy（PDT） of human lymphocytic cells, U-937 in vitro. The PDT efficiency is influenced by the concentration and incubation time. Generally, for ALA and its alkylester methanesulfonates, the cell survival rate decreases and the accumulation ability of PplX increases with the concentration and incubation time. We found that the longer carbon chain methanesulfonates（C5-S, C6-S, C8-S） exhibit better PDT effect than ALA methanesulfonate. This possibly provides a promising route to the clinical application ofPplX-mediated PDT to cancer cell.
文摘Objective To optimize experimental parameters for the photosensitization of 5-aminolevulinic acid (ALA) in promyelocytic leukemia cell HL60 and compare them with normal human peripheral blood mononuclear cell (PBMC). Methods ALA incubation time, wavelength applied to irradiate, concentration of ALA incubated, irradiation fluence may modulate the effect of 5-aminolevulinic acid based Photodynamic Therapy (ALA-PDT).The high-pressure mercury lamps of 400W served as light source, the interference filter of 410nm, 432nm, 545nm, 577nm were used to select the specific wavelength. Fluorescence microscope was used to detect the fluorescence intensity and location of protoporphyrin IX (PpIX) endogenously produced by ALA. MTT assay was used to measure the survival of cell. Flow cytometry with ANNEXIN V FITC kit (contains annexin V FITC, binding buffer and PI) was used to detect the mode of cell death. Results ① 1mmol/L ALA incubated 1×105/mL HL60 cell line for 4 hours, the maximum fluorescence of ALA induced PpIX was detected in cytomembrane. ② Irradiated with 410nm for 14.4J/cm2 can result in the minimum survivability of HL60 cell. ③ The main mode of HL60 cell death caused by ALA-PDT is necrosis. Conclusion ALA for 1mmol/L, 4 hours for dark incubation time, 410nm for irradiation wavelength, 14.4J/cm2 for irradiation fluence were the optimal parameters to selectively eliminate promyelocytic leukemia cell HL60 by ALA based PDT. The photosensitization of ALA based PDT caused the necrosis of HL60 cell, so it could be used for inactivation of certain leukemia cells.
文摘Objective:The aim of the study was to compare the effects of photodynamic therapy(PDT) with δ-aminolevulinic acid(ALA) for patients with different kinds of skin cancers and pre-cancers.Methods:The present study enrolled seventyfive cases,which included 17 cases of actinic keratosis(AK),9 cases of Bowen's disease,11 cases of superficial basal cell carcinomas(BCC),23 cases of nodule basal cell carcinomas and 15 cases of squamous cell carcinomas(SCC),and every patient had single lesion.All patients were treated with 20% ALA topically and He-Ne laser weekly for three times,and followed up 1-3 years.Results:After therapy,the rates of complete reaction(CR) were 100% in AK lesions,77.8% in Bowen's diseases,90.9% in superficial BCCs,47.8% in nodule BCCs,and 50.3% in SCCs,which had significant differences among these five kinds of lesions(H = 18.27,P < 0.05).The therapeutic effectiveness of ALA-PDT for AK was superior to that of Bowen's disease(Q = 4.364,P < 0.05),superficial BCC(Q = 5.55,P < 0.01),SCC(Q = 8.94,P < 0.01) and nodule BCC(Q = 17.91,P < 0.01);the effect of Bowen's disease was better than that of SCC(Q = 7.8,P < 0.01),nodule BCC(Q = 13.44,P < 0.01);the effect of superficial BCC was better than that of SCC(Q = 9.73,P < 0.01),nodule BCC(Q = 16.28,P < 0.01),but similar with Bowen's disease(Q = 0.96,P > 0.05);the effect of SCC was better than that of nodule BCC(Q = 17.74,P < 0.01).Conclusion:Our study shows that therapeutic effectiveness of ALA-PDT for AK is best in five diseases,and Bowen's disease and superficial BCC are secondary,while nodule BCC and SCC are at the bottom.
基金supported by National Natural Science Foundation of China(grant No.81571902,31872742).
文摘Background:Epidermal stem cells(EpSCs)that reside in cutaneous hair follicles and the basal layer of the epidermis are indispensable for wound healing and skin homeostasis.Little is known about the effects of photochemical activation on EpSC differentiation,proliferation and migration during wound healing.The present study aimed to determine the effects of photodynamic therapy(PDT)on wound healing in vivo and in vitro.Methods:We created mouse full-thickness skin resection models and applied 5-aminolevulinic acid(ALA)for PDT to the wound beds.Wound healing was analysed by gross evaluation and haematoxylin–eosin staining in vivo.In cultured EpSCs,protein expression was measured using flow cytometry and immunohistochemistry.Cell migration was examined using a scratch model;apoptosis and differentiation were measured using flow cytometry.Results:PDT accelerated wound closure by enhancing EpSC differentiation,proliferation and migration,thereby promoting re-epithelialization and angiogenesis.PDT inhibited inflammatory infiltration and expression of proinflammatory cytokines,whereas the secretion of growth factors was greater than in other groups.The proportion of transient amplifying cells was significantly greater in vivo and in vitro in the PDT groups.EpSC migration was markedly enhanced after ALAinduced PDT.Conclusions:Topical ALA-induced PDT stimulates wound healing by enhancing re-epithelialization,promoting angiogenesis as well as modulating skin homeostasis.This work provides a preliminary theoretical foundation for the clinical administration of topical ALA-induced PDT in skin wound healing.
