CN109453381A - A kind of low temperature plasma activating fluid inhibiting the growth of malignant tumour of skin cell - Google Patents
A kind of low temperature plasma activating fluid inhibiting the growth of malignant tumour of skin cell Download PDFInfo
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- CN109453381A CN109453381A CN201811230690.4A CN201811230690A CN109453381A CN 109453381 A CN109453381 A CN 109453381A CN 201811230690 A CN201811230690 A CN 201811230690A CN 109453381 A CN109453381 A CN 109453381A
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Abstract
The invention belongs to medical material technical fields, disclose a kind of low temperature plasma (CAP) activating fluid of inhibition malignant tumour of skin cell growth, using cell culture fluid (DMEM) as substrate liquid, DMEM is activated using CAP device, to obtain low temperature plasma activating fluid (CAP-DMEM), it reuses CAP-DMEM and acts on application on human skin basal-cell carcinoma in vivo and in vitro, squamous cell carcinoma and malignant melanoma cell, it was found that the CAP-DMEM property of can choose inhibits the growth of above-mentioned 3 kinds of malignant cells, and inhibit the hyperplasia of tumour after nude mice plantation in vivo.Anti-cancer methods CAP-DMEM of the present invention novel as one bring new hope to future clinical treatment malignant tumour of skin.CAP-DMEM is injected directly into tumour body and is treated, growth of tumour cell can be effectively inhibited, it is not damaged to surrounding normal cell.
Description
Technical field
The invention belongs to medical material technical field more particularly to a kind of low temperature for inhibiting the growth of malignant tumour of skin cell
Plasma activating fluid.
Background technique
Currently, the prior art commonly used in the trade is such that
Plasma is generated after gas is ionized by the molecular ion in electroneutral of positively or negatively grain
Change gas, active material mainly include electronics, negative ions, ground state atom, excited atom, living radical (such as ROS,
RNS etc.) and a small amount of ultraviolet light etc. (UV).Recently as the development of science and technology, low temperature plasma (Cold atmospheric
Plasma, CAP) due to its to biological tissue there are selectively actings simultaneously, not to institute's affected tissues generate thermal damage, gradually by
Pay attention to people.Research finds that CAP jet action can kill its surface microorganism, wound healing in organism surface,
Also with the growth of selective depression tumour cell.But the penetrability of directly CAP jet stream is limited, it is difficult to realize to have to entity tumor
Effect treatment.After research discovery CAP directly acts on specific liquid environment, these can be made by after action of plasma recently
Liquid generates the biological characteristics with similar direct plasma jet stream, can also equally play selective depression tumor proliferative
Effect, this further expansion application potential of the plasma on medical domain.Liquid after being activated by plasma claims
For " plasma activating fluid ".Many researchs confirm that the daughters such as low temperature activating fluid is more than 20 kinds or more to inside and outside in recent years
For malignant tumour there are specific selective depression effect, these tumours include cervical carcinoma, breast cancer, osteosarcoma, colorectal cancer and mind
Through glioma etc..
But the specific mechanism of current low temperature plasma and plasma activating fluid selective depression tumour cell is still unknown
Really, the mechanism of existing research discovery plasma jet and plasma activating fluid selective depression tumour cell is by releasing
Put the comprehensive function result of active oxygen (ROS), active nitrogen (RNS), free radical, ultraviolet light, charged particle and electric field.Research is simultaneously
Show that its bioactivity of the plasma activating fluid of different substrate liquids generations has differences.
In conclusion the prior art malignant tumour of skin treatment presently, there are limitation be:
Malignant tumour of skin mainly has basal-cell carcinoma, squamous cell carcinoma and the big kinds of tumor of chromoma three, close
The disease incidence of year these three malignant tumours is constantly in ascendant trend.Current main treatment method includes operation, radiotherapy and change
Treat etc., but have some limitations to some extent.Many patients there is no effective treatment, three kinds especially above-mentioned
Treatment method is stern challenge for advanced stage and aged patient.The adaptation range ratio of malignant tumour of skin operative treatment
Relatively narrower clinically specifically includes that most of gerontal patient；Constitution is poor to be difficult to be resistant to operator；Cutaneum carcinoma area is larger, invades
Violate the deeper patient of depth；It needs dermatoplasty but the patient etc. for transplanting required skin is provided without sufficient donor site.Though energy of performing the operation
Effectively excision disease damage tissue, but surgery anesthesia and operation itself can induce various complication, and perform the operation to shift advanced stage and suffer from
For person without therapeutic potential, the effect of especially chromoma operative treatment is very limited.Chemotherapy be treat at present it is pernicious
One of main method of cutaneum carcinoma, but can effectively kill the chemotherapeutics of rodent ulcer and squamous cell carcinoma very
It is few, and specificity is not high, while killing tumour cell, there is also toxic side effects, such as chemotherapy often to make for normal tissue cell
At alopecia, Severe gastrointestinal reaction, whole blood trace elements etc., serious person even threatens patient vitals.The chemotherapy of chromoma is made
With also very limited, be only applicable to post-operative adjuvant therapy, should not the Advanced Malignant that shifts extensively of patients undergoing hepatectomy or whole body it is black
The palliative therapy of plain tumor.Radiotherapy is only applicable to the treatment of local tumor, difficult for the malignant melanoma cell of popularity transfer
With treatment, but radiotherapy will cause local tissue necrosis, scar formation and radiodermatitis etc., in treatment cutaneum carcinoma simultaneously to normal
Tissue can also have an impact and limit the application of radiotherapy.
Solve the difficulty and meaning of above-mentioned technical problem:
About the treatment of malignant tumour of skin, there is no high selectivity chemotherapeutics, surgical resection difficulty clinically
Greatly, many patients are difficult to be resistant to operation, and there is also significant limitations for radiotherapy.
In conclusion low temperature plasma activating fluid is proved up to more than 20 human malignancies to culture in vitro
The selective inhibition growth effects of cell, using plasma jet stream of the present invention handle malignant tumour of skin cell in vitro and also send out
Now the property of can choose inhibits the growth of malignant tumour of skin cell；But in view of Skin Tumor Cells it is in entity growth, directly
It is limited to connect plasma jet penetration depth, after the present invention acts on cell culture medium using plasma jet, obtain etc. from
Daughter activating fluid (CAP-DMEM), then Skin Tumor Cells are acted in vitro, discovery CAP-DMEM also being capable of selective depression
Rodent ulcer, squamous cell carcinoma and malignant melanoma cell, and confirmed in transplanted tumor in nude mice in vitro.
The meaning that using plasma activating fluid handles tumour is, overcomes the office of plasma jet penetration depth
It is sex-limited, plasma activating fluid can be injected directly into knurl and be treated.
Using plasma activating fluid not only overcomes the problem of penetration depth limitation, can also be plasma activating fluid
It is injected directly into internal a variety of solid tumor bodies and is treated, be similar to Local chemiotherapy.But because to normal group
Not the problem of knitting cell does not have depression effect, latent lesion histiocytic so as to avoid similar Chemotherapeutic Drugs On Normal.
Summary of the invention
In view of the problems of the existing technology, the present invention provides a kind of low temperature of inhibition malignant tumour of skin cell growth
Plasma activating fluid.
The invention is realized in this way a kind of low temperature plasma activating fluid for inhibiting the growth of malignant tumour of skin cell
Preparation method, comprising:
Utilize the sterile inoculum of corona treatment caused by low-temperature plasma device (DMEM) 120s-
300s is obtained low temperature plasma activating fluid (CAP-DMEM)；DMEM (dulbecco's modifiedeagle medium),
It is a kind of culture medium containing various amino acid and glucose, is widely used in production of vaccine and various virus host cells primary
Cell culture and single cell culture.
Further, the preparation method packet of the low temperature plasma activating fluid for inhibiting the growth of malignant tumour of skin cell
It includes:
The ground wire of low-temperature plasma device is inserted into sterile DMEM, after powering on, is launched by thin copper bar
Gas ions jet stream, plasma jet obtain low temperature plasma activating fluid CAP-DMEM after handling 120s.
Further, the preparation method packet of the low temperature plasma activating fluid for inhibiting the growth of malignant tumour of skin cell
It includes: the ground wire of low-temperature plasma device being inserted into sterile DMEM, after powering on, thin copper bar can launch plasma and penetrate
Stream, plasma jet obtain low temperature plasma activating fluid CAP-DMEM after handling 240s.
Further, the preparation method packet of the low temperature plasma activating fluid for inhibiting the growth of malignant tumour of skin cell
It includes: the ground wire of low-temperature plasma device being inserted into sterile DMEM, after powering on, thin copper bar is launched plasma and penetrated
Stream, plasma jet obtain low temperature plasma activating fluid: CAP-DMEM after handling 300s.
Another object of the present invention is to provide a kind of low temperature plasmas for inhibiting cervical carcinoma malignant cell growth
Activating fluid.
Another object of the present invention is to provide a kind of low temperature plasmas for inhibiting breast cancer malignant cell growth
Activating fluid.
Another object of the present invention is to provide a kind of low temperature plasmas for inhibiting osteosarcoma malignant cell growth
Activating fluid.
Another object of the present invention is to provide a kind of low temperature plasmas for inhibiting colorectal cancer malignant cell growth
Activating fluid.
Another object of the present invention is to provide a kind of low temperature etc. for inhibiting glioma malignant cell growth from
Daughter activating fluid.
In conclusion advantages of the present invention and good effect are as follows:
It is of the invention innovative using DMEM as substrate liquid, DMEM is activated using low-temperature plasma device, to obtain
CAP-DMEM is obtained, CAP-DMEM is reused and acts on application on human skin basal-cell carcinoma, squamous cell carcinoma and pernicious melanocyte in vivo and in vitro
Oncocyte, the discovery CAP-DMEM property of can choose inhibits the growth of above-mentioned 3 kinds of malignant cells, and inhibits nude mice in vivo
The hyperplasia of tumour after plantation.After CAP-DMEM handles Skin Tumor Cells, with the increasing of the CAP-DMEM jet stream time received
Add, Skin Tumor Cells volume gradually becomes smaller, and cell gradually loses normal morphology and starts to be rounded, and nucleus starts pyknosis, largely
Cell falls off from culture dish, as shown in Figure 1.It is as shown in Figure 2 and Figure 3 for cell viability after CAP-DMEM processing and apoptosis degree.
CAP-DMEM shows apparent dose dependent and time dependence to the metabolic activity of application on human skin epidermoid carcinoma cell, when
When DMEM receives the plasma jet time and reaches 120s, the metabolic activity of Skin Squamous Cell Carcinoma, which has shown that, apparent to be inhibited to make
With.When the jet stream time reaching 300s, about 80% or so cell has lost vitality (Fig. 2).Meanwhile CAP-DMEM inhibits cell
Metabolic activity with it caused by it is apparent apoptosis-related, CAP-DMEM apoptosis caused by malignant tumour of skin also shows that
Apparent dose dependent (Fig. 3).
Therefore, the low temperature plasma activating fluid anti-cancer methods novel as one are swollen to future clinical treatment cutaneous malignant
Tumor brings new hope.It can be treated, can effectively be inhibited by the way that CAP-DMEM is injected directly into tumour body
Growth of tumour cell, it is not damaged to surrounding normal cell.
Its advantage is that surgical operation, radiation and chemotherapy can be overcome to state the limitation in 3 kinds of malignant cutaneous cancers in the treatment,
Can effectively tissue tumor cells growth while, it is not damaged to surrounding normal skin histology, therefore locally injecting can be passed through
Achieve the purpose that the above-mentioned 3 kinds of tumours for the treatment of, without considering that bad hair brought by operation, chemotherapy and radiation is answered.
Detailed description of the invention
Fig. 1 is Skin Tumor Cells metamorphosis figure after CAP-DMEM processing provided in an embodiment of the present invention.
Fig. 2 is that mtt assay provided in an embodiment of the present invention surveys suppression of the CAP-DMEM to malignant tumour of skin cell metabolism vigor
Action diagram processed.
The bis- dye method CAP-DMEM of Fig. 3 AnnexinV/PI generate apoptotic effect figure to Skin Tumor Cells.
Fig. 4 is the low temperature plasma activating fluid of inhibition malignant tumour of skin cell growth provided in an embodiment of the present invention
Preparation method flow chart.
Fig. 5 is low temperature plasma excitation apparatus schematic diagram provided in an embodiment of the present invention.
In figure: 1, resistance；2, thin copper bar composite structure；3, low temperature plasma；4, dielectric material；5, CAP-DMEM low temperature
Plasma activating fluid；6, DC power supply.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
At present low temperature plasma and plasma activating fluid it is selectively antitumor specific mechanism it is still indefinite, clinically
It is used only for treatment basal-cell carcinoma, squamous cell carcinoma and chromoma.
Fig. 1 is Skin Tumor Cells metamorphosis figure after CAP-DMEM processing provided in an embodiment of the present invention.
Fig. 2 is that mtt assay provided in an embodiment of the present invention surveys suppression of the CAP-DMEM to malignant tumour of skin cell metabolism vigor
Action diagram processed.
The bis- dye method CAP-DMEM of Fig. 3 AnnexinV/PI generate apoptotic effect figure to Skin Tumor Cells.
Such as Fig. 4, the low temperature plasma activating fluid provided in an embodiment of the present invention for inhibiting the growth of malignant tumour of skin cell
The preparation method comprises the following steps:
The ground wire of low-temperature plasma device is inserted into sterile DMEM, after powering on, thin copper bar can be launched etc. from
Daughter jet stream；
Sterile DMEM appropriate time (120s- is handled using plasma jet caused by low-temperature plasma device
300s), DMEM, i.e. CAP-DMEM after obtaining low temperature plasma activation.
Below with reference to low-temperature plasma device, the invention will be further described.
The present invention handles sterile DMEM using low-temperature plasma device, thus after obtaining low temperature plasma activation
DMEM, i.e. CAP-DMEM.The following are main device and implementation steps:
1) low temperature plasma excitation apparatus
The low temperature plasma excitation apparatus that the invention uses by a cylindrical polytetrafluoroethylene (PTFE) bucket as insulation crust,
Internal parallel places several professional thin copper bars (diameter 1mm) as lattice electrode and connects direct current high voltage pulses power supply, high-voltage line
An appropriately sized resistance is connected, for limiting electric discharge.(being illustrated in fig. 5 shown below)
In figure, low temperature plasma excitation apparatus includes: resistance 1, thin copper bar composite structure 2, low temperature plasma 3, is situated between
Electric material 4, CAP-DMEM low temperature plasma activating fluid 5, DC power supply 6.
2) obtains low temperature plasma activating fluid step:
Ground wire is inserted into sterile DMEM, after powering on, thin copper bar can launch plasma jet, and plasma is penetrated
It can be obtained low temperature plasma activating fluid: CAP-DMEM after stream process DMEM appropriate time.
The present invention is made combined with specific embodiments below further.
Embodiment 1
1) ground wire of low-temperature plasma device is inserted into sterile DMEM, after powering on, thin copper bar can be launched
Gas ions jet stream, plasma jet can be obtained low temperature plasma activating fluid: CAP-DMEM (120s) after handling 120s.
2) ground wire of low-temperature plasma device is inserted into sterile DMEM, after powering on, thin copper bar can be launched
Gas ions jet stream, plasma jet can be obtained low temperature plasma activating fluid: CAP-DMEM (240s) after handling 240s.
3) ground wire of low-temperature plasma device is inserted into sterile DMEM, after powering on, thin copper bar can be launched
Gas ions jet stream, plasma jet can be obtained low temperature plasma activating fluid: CAP-DMEM (300s) after handling 300s.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (9)
1. a kind of preparation method for the low temperature plasma activating fluid for inhibiting the growth of malignant tumour of skin cell, which is characterized in that
The preparation method of low temperature plasma activating fluid for inhibiting the growth of malignant tumour of skin cell includes:
Using the sterile inoculum DMEM120s-300s of corona treatment caused by low-temperature plasma device, obtain
Low temperature plasma activating fluid CAP-DMEM.
2. inhibiting the preparation side of the low temperature plasma activating fluid of malignant tumour of skin cell growth as described in claim 1
Method, which is characterized in that the preparation method of low temperature plasma activating fluid for inhibiting the growth of malignant tumour of skin cell includes:
The ground wire of low-temperature plasma device is inserted into sterile DMEM, after powering on, thin copper bar is launched plasma and penetrated
Stream, plasma light beam obtain low temperature plasma activating fluid CAP-DMEM after irradiating 120s.
3. inhibiting the preparation side of the low temperature plasma activating fluid of malignant tumour of skin cell growth as described in claim 1
Method, which is characterized in that the preparation method of low temperature plasma activating fluid for inhibiting the growth of malignant tumour of skin cell includes:
The ground wire of low-temperature plasma device is inserted into sterile DMEM, after powering on, thin copper bar can launch plasma jet,
Plasma light beam obtains low temperature plasma activating fluid CAP-DMEM after irradiating 240s.
4. inhibiting the preparation side of the low temperature plasma activating fluid of malignant tumour of skin cell growth as described in claim 1
Method, which is characterized in that the preparation method of low temperature plasma activating fluid for inhibiting the growth of malignant tumour of skin cell includes:
The ground wire of low-temperature plasma device is inserted into sterile DMEM, after powering on, thin copper bar launches plasma jet, etc.
Gas ions light beam obtains low temperature plasma activating fluid: CAP-DMEM after irradiating 300s.
5. a kind of low temperature plasma using inhibition malignant tumour of skin cell growth described in 4 any one of Claims 1 to 4
The low temperature plasma activating fluid of the inhibition uterine neck malignant cell growth of the preparation method preparation of activating fluid.
6. a kind of low temperature plasma using inhibition malignant tumour of skin cell growth described in 4 any one of Claims 1 to 4
The low temperature plasma activating fluid of the inhibition breast cancer malignant cell growth of the preparation method preparation of activating fluid.
7. a kind of low temperature plasma using inhibition malignant tumour of skin cell growth described in 4 any one of Claims 1 to 4
The low temperature plasma activating fluid of the inhibition osteosarcoma malignant cell growth of the preparation method preparation of activating fluid.
8. a kind of low temperature plasma using inhibition malignant tumour of skin cell growth described in 4 any one of Claims 1 to 4
The low temperature plasma activating fluid of the inhibition colorectal cancer malignant cell growth of the preparation method preparation of activating fluid.
9. a kind of low temperature plasma using inhibition malignant tumour of skin cell growth described in 4 any one of Claims 1 to 4
The low temperature plasma activating fluid of the inhibition glioma malignant cell growth of the preparation method preparation of activating fluid.
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