Nonlinear optical photodynamic therapy (NLO-PDT) of the cornea
The embodiments include method of nonlinear optical photodynamic therapy of tissue including the steps of providing pulsed infrared laser light for two-photon excited fluorescence tissue exposure, and selectively focusing the pulsed infrared laser light within the tissue at a focal plane to activate a photosensitizing agent to generate free radicals within a highly resolved axial and lateral spatial domain in the tissue. The invention is also directed to an apparatus for performing nonlinear optical photodynamic therapy of tissue including a pulsed infrared laser for providing two-photon excited fluorescence beam tissue exposure, a scanner for selectively and controllably moving the tissue and the beam relative to each other, and optics for selectively focusing the pulsed infrared laser light within the tissue at a point in a focal plane to activate a photosensitizing agent to generate free radicals within a highly resolved axial and lateral spatial domain in the tissue.
1. A method of nonlinear optical photodynamic therapy of a cornea, the method comprising:
pretreating the cornea with a photosensitive agent, which is capable of generating free radicals within a treatment volume of the cornea upon laser irradiation; and providing the photodynamic therapy within the treatment volume of the cornea by:
directing to said cornea laser irradiation by a pulsed near-infrared laser light, which provides a multiphoton beam at a predetermined intensity and having a near-infrared wavelength through a focusing objective lens of an apparatus, the multiphoton beam having a beam width and the focusing objective lens having a fixed numerical aperture;
generating a focal volume with the multiphoton beam at a depth in the cornea by reducing an effective numerical aperture of the apparatus, compared to the numerical aperture of the focusing objective lens, to a final value between 0.16 and 0.08 by expanding the beam width impinging on the focusing objective lens, wherein the focal volume comprises is between 150 and 2500 femtoliters; and
moving the focal volume in three dimensions relative to the tissue within a predetermined clinical time span, to define the treatment volume, which is larger than the focal volume.
2. The method of claim 1 , wherein the photosensitive agent comprises riboflavin.
3. The method of claim 1 , wherein the wavelength is 700-960 nm.
4. The method of claim 1 , wherein the pulsed near infrared laser light within the cornea provides an intensity and length of irradiation to cause collagen crosslinking (CXL).
5. The method of claim 1 , wherein the pulsed near infrared laser light within the cornea provides an intensity and length of irradiation that provides anti-microbial mediation.
6. The method of claim 1 , wherein the pulsed near-infrared laser light within the cornea provides an intensity and length of irradiation that inhibits corneal swelling in bullous keratopathy.
7. The method of claim 1 , wherein the pulsed near-infrared laser light within the cornea provides an intensity and length of irradiation that kills cells, bacteria, tumors or neovascular vessels growing into the cornea.
8. The method of claim 1 comprising activating the photosensitizing agent only at a focal plane of the focal volume by limiting intensity and length of irradiation by the pulsed near-infrared laser light within the cornea.
9. The method of claim 1 comprising crosslinking collagen in the cornea by an intensity and length of irradiation by the pulsed near-infrared laser light to precisely stiffen the cornea, wherein the cornea is initially weakened prior to being stiffened.
10. The method of claim 1 comprising adjusting the pulsed near-infrared laser light within the cornea to provide an intensity and length of irradiation that causes corneal stiffening, flattening and steepening to precisely stiffen, flatten and steepen regions of the cornea, thereby treating an astigmatism or a refractive error associated with myopia, hyperopia or presbyopia.
11. The method of claim 1 , wherein the pulsed near infrared laser light within the cornea provides an intensity and length of irradiation that treats bacterial, fungal, or amoebic infections of the cornea without antibiotics.
12. The method of claim 1 , wherein the pulsed near-infrared laser light within the cornea provides an intensity and length of irradiation that kills labeled tumor cells in the cornea following loading with photosensitizing dyes.
13. The method of claim 1 , comprising adjusting an intensity and length of irradiation of the pulsed near-infrared laser light within the cornea to treat a clinical disease.
14. The method of claim 9 comprising treating a keratoconus or a post-LASIK ectasia.
15. The method according to claim 13 , wherein the clinical disease is selected from the group consisting of keratoconus, post-LASIK ectasia, astigmatism, myopia, hyperopia, presbyopia, infection and an ocular tumor.