In vivo kinetics and spectra of 5-aminolaevulinic acid-induced fluorescence in an amelanotic melanoma of the hamster.

For successful photodynamic diagnosis (PDD) and effective photodynamic therapy (PDT) with the clinically used 'photosensitiser' 5-aminolaevulinic acid (ALA), knowledge of the maximal fluorescence intensity and of the maximal tumour-host tissue fluorescence ratio following systemic or local application is required. Therefore, time course and type of porphyrin accumulation were investigated in neoplastic and surrounding host tissue by measuring the kinetics and spectra of ALA-induced fluorescence in vivo. Experiments were performed in the amelanotic melanoma A-Mel-3 grown in the dorsal skinfold chamber preparation of Syrian golden hamsters. The kinetics of fluorescent porphyrins was quantified up to 24 h after i.v. injection of 100 mg kg-1, 500 mg kg-1 or 1,000 mg kg-1 body weight ALA by intravital fluorescence microscopy and digital image analysis (n = 18). In separate experiments fluorescence spectra were obtained for each dose by a simultaneous optical multichannel analysing device (n = 3). A three-compartment model was developed to simulate fluorescence kinetics in tumours. Maximal fluorescence intensity (per cent of reference standard; mean +/- s.e.) in the tumour arose 150 min post injection (p.i.) (1,000 mg kg-1, 109 +/- 34%; 500 mg kg-1, 148 +/- 36%) and 120 min p.i. (100 mg kg-1, 16 +/- 8%). The fluorescence in the surrounding host tissue was far less and reached its maximum at 240 min (100 mg kg-1, 6 +/- 3%) and 360 min p.i. (500 mg kg-1, 50 +/- 8%) and (1,000 mg kg-1, 6 +/- 19%). Maximal tumour-host tissue ratio (90:1) was encountered at 90 min after injection of 500 mg kg-1. The spectra of tissue fluorescence showed maxima at 637 nm and 704 nm respectively. After 300 min (host tissue) and 360 min (tumour tissue) additional emission bands at 618 nm and 678 nm were detected. These bands indicate the presence of protoporphyrin IX (PPIX) and of another porphyrin species in the tumour not identified yet. Tumour selectivity of ALA-induced PPIX accumulation occurs only during a distinct interval depending on the administered dose. Based on the presented data the optimal time for PDD and PDT in this model following intravenous administration of 500 mg kg-1 ALA would be around 90 min and 150 min respectively. The transient selectivity is probably caused by an earlier and higher uptake of ALA in the neoplastic tissue most likely as a result of increased vascular permeability of tumours as supported by the mathematical model.

[1]  Herbert Stepp,et al.  Delta-ALA-assisted fluorescence detection of cancer in the urinary bladder , 1994, Other Conferences.

[2]  S. G. Bown,et al.  Photodynamic therapy of the normal rat stomach: a comparative study between di-sulphonated aluminium phthalocyanine and 5-aminolaevulinic acid. , 1992, British Journal of Cancer.

[3]  G. Kuhnle,et al.  Relation between autoradiographically measured blood flow and ATP concentrations obtained from imaging bioluminescence in tumors following hyperthermia , 1993, International journal of cancer.

[4]  D. Brault Physical chemistry of porphyrins and their interactions with membranes: the importance of pH. , 1990, Journal of photochemistry and photobiology. B, Biology.

[5]  D. Phillips,et al.  Fluorescence distribution and photodynamic effect of ALA-induced PP IX in the DMH rat colonic tumour model. , 1992, British Journal of Cancer.

[6]  H. Kerl,et al.  Photodynamic therapy in patient with xeroderma pigmentosum , 1991, The Lancet.

[7]  A. Batlle,et al.  Porphyrins, porphyrias, cancer and photodynamic therapy--a model for carcinogenesis. , 1993, Journal of photochemistry and photobiology. B, Biology.

[8]  K Messmer,et al.  Quantitative analysis of microvascular structure and function in the amelanotic melanoma A-Mel-3. , 1981, Cancer research.

[9]  F. Berr,et al.  Effect of dietary n-3 versus n-6 polyunsaturated fatty acids on hepatic excretion of cholesterol in the hamster. , 1993, Journal of lipid research.

[10]  J C Kennedy,et al.  NON‐INVASIVE TECHNIQUE FOR OBTAINING FLUORESCENCE EXCITATION AND EMISSION SPECTRA IN VIVO , 1986, Photochemistry and photobiology.

[11]  J Moan,et al.  Distribution and photosensitizing efficiency of porphyrins induced by application of exogenous 5‐aminolevulinic acid in mice bearing mammary carcinoma , 1992, International journal of cancer.

[12]  R K Jain,et al.  Extravascular diffusion in normal and neoplastic tissues. , 1984, Cancer research.

[13]  J. Zenklusen,et al.  Influence of hepatic tumors caused by diethylnitrosamine on hexachlorobenzene-induced porphyria in rats. , 1991, Cancer letters.

[14]  H. Dailey,et al.  Differential interaction of porphyrins used in photoradiation therapy with ferrochelatase. , 1984, The Biochemical journal.

[15]  D. Vernon,et al.  Endogenous porphyrin distribution induced by 5-aminolaevulinic acid in the tissue layers of the gastrointestinal tract. , 1993, Journal of photochemistry and photobiology. B, Biology.

[16]  N Kollias,et al.  Effects of photodynamic therapy with topical application of 5-aminolevulinic acid on normal skin of hairless guinea pigs. , 1992, Journal of photochemistry and photobiology. B, Biology.

[17]  R K Jain,et al.  Therapeutic implications of tumor physiology , 1991, Current opinion in oncology.

[18]  P. Speight,et al.  Photodynamic therapy of oral cancer: photosensitisation with systemic aminolaevulinic acid , 1993, The Lancet.

[19]  R K Jain,et al.  Transport of molecules in the tumor interstitium: a review. , 1987, Cancer research.

[20]  J C Kennedy,et al.  Endogenous protoporphyrin IX, a clinically useful photosensitizer for photodynamic therapy. , 1992, Journal of photochemistry and photobiology. B, Biology.

[21]  R. van Hillegersberg,et al.  Selective accumulation of endogenously produced porphyrins in a liver metastasis model in rats. , 1992, Gastroenterology.

[22]  J. Kennedy,et al.  Photodynamic therapy with endogenous protoporphyrin IX: basic principles and present clinical experience. , 1990, Journal of photochemistry and photobiology. B, Biology.

[23]  D. Jocham,et al.  Tumour localisation kinetics of photofrin and three synthetic porphyrinoids in an amelanotic melanoma of the hamster. , 1993, British Journal of Cancer.

[24]  P. Okunieff,et al.  Blood flow, oxygen and nutrient supply, and metabolic microenvironment of human tumors: a review. , 1989, Cancer research.

[25]  N. Navone,et al.  Heme biosynthesis in human breast cancer--mimetic "in vitro" studies and some heme enzymic activity levels. , 1990, The International journal of biochemistry.

[26]  J. Kennedy,et al.  Experimental Porphyric Neuropathy: A Preliminary Report , 1981, Canadian Journal of Neurological Sciences / Journal Canadien des Sciences Neurologiques.

[27]  J. Reynoldson,et al.  Neuropharmacology of delta-aminolaevulinic acid—I Effect of acute administration in rodents , 1984, Neuropharmacology.

[28]  P. Gullino The internal milieu of tumors. , 1966, Progress in experimental tumor research.

[29]  S. Arkins,et al.  PHOTODESTRUCTION OF TUMOR CELLS BY INDUCTION OF ENDOGENOUS ACCUMULATION OF PROTOPORPHYRIN IX: ENHANCEMENT BY 1, 10‐PHENANTHROLINE , 1992, Photochemistry and photobiology.

[30]  F. Becker,et al.  Heme synthesis in normal mouse liver and mouse liver tumors. , 1990, Cancer research.

[31]  J C Kennedy,et al.  Phototoxic damage to sebaceous glands and hair follicles of mice after systemic administration of 5-aminolevulinic acid correlates with localized protoporphyrin IX fluorescence. , 1990, The American journal of pathology.

[32]  R. Jain,et al.  Microvascular permeability of normal and neoplastic tissues. , 1986, Microvascular research.

[33]  W. Durán,et al.  Experimental determination of the linear correlation between in vivo TV fluorescence intensity and vascular and tissue FITC-DX concentrations. , 1991, Microvascular research.

[34]  S. Ida,et al.  Excretion of porphyrins in urine and bile after the administration of delta-aminolevulinic acid. , 1978, The Journal of laboratory and clinical medicine.

[35]  T. Dougherty Optical Methods for Tumor Treatment and Detection: Mechanisms and Techniques in Photodynamic Therapy XI , 1994 .

[36]  K. Messmer,et al.  Technical report—a new chamber technique for microvascular studies in unanesthetized hamsters , 1980, Research in experimental medicine. Zeitschrift fur die gesamte experimentelle Medizin einschliesslich experimenteller Chirurgie.

[37]  H Kerl,et al.  Topical photodynamic therapy with endogenous porphyrins after application of 5-aminolevulinic acid. An alternative treatment modality for solar keratoses, superficial squamous cell carcinomas, and basal cell carcinomas? , 1993, Journal of the American Academy of Dermatology.

[38]  M Intaglietta,et al.  Tissue perfusion inhomogeneity during early tumor growth in rats. , 1979, Journal of the National Cancer Institute.

[39]  M Landthaler,et al.  PENETRATION POTENCY OF TOPICAL APPLIED δ‐AMINOLEVULINIC ACID FOR PHOTODYNAMIC THERAPY OF BASAL CELL CARCINOMA * , 1994, Photochemistry and photobiology.

[40]  A. Neuberger,et al.  The metabolism of delta -aminolaevulic acid. 2. Normal pathways, studied with the aid of 14C. , 1956, The Biochemical journal.

[41]  G. Schrodt,et al.  Transplantable tumors of the Syrian (golden) hamster. I. Tumors of the alimentary tract, endocrine glands and melanomas. , 1961, Cancer research.